def test_banding_to_disk(ksize, memory, numbands):
"""
Test accuracy of banding in terms of the data structure contents.
Stronger than the functional in-memory test, this function tests whether
a computing k-mer abundances in banding mode produces the same data
structure as counting k-mer abundances in the normal fashion.
"""
infile = utils.get_test_data('banding-reads.fq')
path1 = utils.get_temp_filename('normal.ct')
path2 = utils.get_temp_filename('banding.ct')
ct = khmer.Counttable(ksize, memory / 4, 4)
ct.consume_seqfile(infile)
ct.save(path1)
fpr = khmer.calc_expected_collisions(ct)
print('FPR', fpr)
ct = khmer.Counttable(ksize, memory / 4, 4)
for band in range(numbands):
ct.consume_seqfile_banding(infile, numbands, band)
ct.save(path2)
fpr = khmer.calc_expected_collisions(ct)
print('FPR', fpr)
with open(path1, 'rb') as f1, open(path2, 'rb') as f2:
assert f1.read() == f2.read()
def test_banding_to_disk(ksize, memory, numbands):
"""
Test accuracy of banding in terms of the data structure contents.
Stronger than the functional in-memory test, this function tests whether
a computing k-mer abundances in banding mode produces the same data
structure as counting k-mer abundances in the normal fashion.
"""
infile = utils.get_test_data('banding-reads.fq')
path1 = utils.get_temp_filename('normal.ct')
path2 = utils.get_temp_filename('banding.ct')
ct = khmer.Counttable(ksize, memory / 4, 4)
ct.consume_seqfile(infile)
ct.save(path1)
fpr = khmer.calc_expected_collisions(ct)
print('FPR', fpr)
ct = khmer.Counttable(ksize, memory / 4, 4)
for band in range(numbands):
ct.consume_seqfile_banding(infile, numbands, band)
ct.save(path2)
fpr = khmer.calc_expected_collisions(ct)
print('FPR', fpr)
with open(path1, 'rb') as f1, open(path2, 'rb') as f2:
assert f1.read() == f2.read()
def normalizeByMedian(cf):
"""Wrapper for the normalizeByMedian_impl function."""
kmersize = cf.get_parameter('kmersize', 'int')
minhashsize = cf.get_parameter('minhashsize', 'float')
nhashes = cf.get_parameter('nhashes', 'int')
cutoff = cf.get_parameter('cutoff', 'int')
inputfile = cf.get_input('inputfile')
outputfile = cf.get_output('outputfile')
infp = screed.open(inputfile)
outfp = open(outputfile, 'w')
discarded, total, ht, n = normalizeByMedian_impl(cf, infp, outfp, kmersize, \
minhashsize, nhashes, cutoff)
outfp.close()
infp.close()
if -1 < n:
percent_kept = int(100. - discarded / float(total) * 100.)
cf.write_log("DONE with %s; kept %s of %s or %s%%" % (inputfile, \
total - discarded, total, percent_kept))
cf.write_log("Output in %s" % outputfile)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
cf.write_log("fp rate estimated to be %1.3f" % fp_rate)
if fp_rate > 0.20:
cf.write_error("ERROR: the counting hash is too small.")
cf.write_error("Increase the hashsize/num ht.")
return constants.GENERIC_ERROR
return constants.OK
def __str__(self):
return ('**Leaf:{name} [occupied: {nb}, fpr: {fpr:.2}]'
'-> {metadata}'.format(name=self.name,
metadata=self.metadata,
nb=self.data.n_occupied(),
fpr=khmer.calc_expected_collisions(
self.data, True, 1.1)))
def __str__(self):
return "**Leaf:{name} [occupied: {nb}, fpr: {fpr:.2}] -> {metadata}".format(
name=self.name,
metadata=self.metadata,
nb=self.graph.n_occupied(),
fpr=khmer.calc_expected_collisions(self.graph, True, 1.1),
)
def main():
info("filter-abund-single.py", ["counting"])
args = get_parser().parse_args()
check_file_status(args.datafile)
check_space([args.datafile])
if args.savetable:
check_space_for_hashtable(args.n_tables * args.min_tablesize)
report_on_config(args)
config = khmer.get_config()
config.set_reads_input_buffer_size(args.threads * 64 * 1024)
print "making k-mer counting table"
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize, args.n_tables, args.threads)
# first, load reads into hash table
rparser = khmer.ReadParser(args.datafile, args.threads)
threads = []
print "consuming input, round 1 --", args.datafile
for _ in xrange(args.threads):
cur_thread = threading.Thread(target=htable.consume_fasta_with_reads_parser, args=(rparser,))
threads.append(cur_thread)
cur_thread.start()
for _ in threads:
_.join()
fp_rate = khmer.calc_expected_collisions(htable)
print "fp rate estimated to be %1.3f" % fp_rate
# now, trim.
# the filtering function.
def process_fn(record):
name = record["name"]
seq = record["sequence"]
if "N" in seq:
return None, None
trim_seq, trim_at = htable.trim_on_abundance(seq, args.cutoff)
if trim_at >= args.ksize:
return name, trim_seq
return None, None
# the filtering loop
print "filtering", args.datafile
outfile = os.path.basename(args.datafile) + ".abundfilt"
outfp = open(outfile, "w")
tsp = ThreadedSequenceProcessor(process_fn)
tsp.start(verbose_loader(args.datafile), outfp)
print "output in", outfile
if args.savetable:
print "Saving k-mer counting table filename", args.savetable
print "...saving to", args.savetable
htable.save(args.savetable)
def main():
args = sanitize_help(get_parser()).parse_args()
configure_logging(args.quiet)
check_input_files(args.datafile, args.force)
check_space([args.datafile], args.force)
if args.savegraph:
tablesize = calculate_graphsize(args, "countgraph")
check_space_for_graph(args.savegraph, tablesize, args.force)
report_on_config(args)
log_info("making countgraph")
graph = khmer_args.create_countgraph(args)
# first, load reads into graph
rparser = khmer.ReadParser(args.datafile)
threads = []
log_info("consuming input, round 1 -- {datafile}", datafile=args.datafile)
for _ in range(args.threads):
cur_thread = threading.Thread(target=graph.consume_fasta_with_reads_parser, args=(rparser,))
threads.append(cur_thread)
cur_thread.start()
for _ in threads:
_.join()
log_info("Total number of unique k-mers: {nk}", nk=graph.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(graph, args.force)
log_info("fp rate estimated to be {fpr:1.3f}", fpr=fp_rate)
# the filtering loop
log_info("filtering {datafile}", datafile=args.datafile)
if args.outfile is None:
outfile = os.path.basename(args.datafile) + ".abundfilt"
else:
outfile = args.outfile
outfp = open(outfile, "wb")
outfp = get_file_writer(outfp, args.gzip, args.bzip)
paired_iter = broken_paired_reader(ReadParser(args.datafile), min_length=graph.ksize(), force_single=True)
for n, is_pair, read1, read2 in paired_iter:
assert not is_pair
assert read2 is None
trimmed_record, _ = trim_record(graph, read1, args.cutoff, args.variable_coverage, args.normalize_to)
if trimmed_record:
print((trimmed_record,))
write_record(trimmed_record, outfp)
log_info("output in {outfile}", outfile=outfile)
if args.savegraph:
log_info("Saving k-mer countgraph filename {graph}", graph=args.savegraph)
graph.save(args.savegraph)
def main(args):
info('build-graph.py', ['graph', 'SeqAn'])
report_on_config(args, hashtype='nodegraph')
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
# if optimization args are given, do optimization
args = functions.do_sanity_checking(args, 0.01)
check_space(args.input_filenames, args.force)
check_space_for_hashtable(args, 'nodegraph', args.force)
print('Saving k-mer presence table to %s' % base, file=sys.stderr)
print('Loading kmers from sequences in %s' %
repr(filenames), file=sys.stderr)
if args.no_build_tagset:
print('We WILL NOT build the tagset.', file=sys.stderr)
else:
print('We WILL build the tagset (for partitioning/traversal).',
file=sys.stderr)
print('making nodegraph', file=sys.stderr)
htable = khmer_args.create_nodegraph(args)
functions.build_graph(filenames, htable, args.threads,
not args.no_build_tagset)
print('Total number of unique k-mers: {0}'.format(htable.n_unique_kmers()),
file=sys.stderr)
print('saving k-mer presence table in', base + '.pt', file=sys.stderr)
htable.save(base + '.pt')
if not args.no_build_tagset:
print('saving tagset in', base + '.tagset', file=sys.stderr)
htable.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % htable.n_unique_kmers())
fp_rate = \
khmer.calc_expected_collisions(htable, args.force, max_false_pos=.15)
# 0.18 is ACTUAL MAX. Do not change.
print('false positive rate estimated to be %1.3f' % fp_rate,
file=sys.stderr)
print('\nfalse positive rate estimated to be %1.3f' % fp_rate,
file=info_fp)
print('wrote to', base + '.info and', base + '.pt', file=sys.stderr)
if not args.no_build_tagset:
print('and ' + base + '.tagset', file=sys.stderr)
sys.exit(0)
def main(args):
graph_type = 'nodegraph'
report_on_config(args, graphtype=graph_type)
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
graphsize = calculate_graphsize(args, graph_type)
space_needed = (args.n_tables * graphsize /
khmer._buckets_per_byte[graph_type])
check_space_for_graph(args.output_filename, space_needed, args.force)
print('Saving k-mer nodegraph to %s' % base, file=sys.stderr)
print('Loading kmers from sequences in %s' % repr(filenames),
file=sys.stderr)
if args.no_build_tagset:
print('We WILL NOT build the tagset.', file=sys.stderr)
else:
print('We WILL build the tagset (for partitioning/traversal).',
file=sys.stderr)
print('making nodegraph', file=sys.stderr)
nodegraph = khmer_args.create_nodegraph(args)
oxfuncs.build_graph(filenames, nodegraph, args.threads,
not args.no_build_tagset)
print('Total number of unique k-mers: {0}'.format(
nodegraph.n_unique_kmers()),
file=sys.stderr)
print('saving k-mer nodegraph in', base, file=sys.stderr)
nodegraph.save(base)
if not args.no_build_tagset:
print('saving tagset in', base + '.tagset', file=sys.stderr)
nodegraph.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % nodegraph.n_unique_kmers())
fp_rate = \
khmer.calc_expected_collisions(
nodegraph, args.force, max_false_pos=.15)
# 0.18 is ACTUAL MAX. Do not change.
print('false positive rate estimated to be %1.3f' % fp_rate,
file=sys.stderr)
print('\nfalse positive rate estimated to be %1.3f' % fp_rate,
file=info_fp)
print('wrote to ' + base + '.info and ' + base, file=sys.stderr)
if not args.no_build_tagset:
print('and ' + base + '.tagset', file=sys.stderr)
sys.exit(0)
def main(args):
graph_type = 'nodegraph'
report_on_config(args, graphtype=graph_type)
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
graphsize = calculate_graphsize(args, graph_type)
space_needed = (args.n_tables * graphsize /
khmer._buckets_per_byte[graph_type])
check_space_for_graph(args.output_filename, space_needed, args.force)
print('Saving k-mer nodegraph to %s' % base, file=sys.stderr)
print('Loading kmers from sequences in %s' %
repr(filenames), file=sys.stderr)
if args.no_build_tagset:
print('We WILL NOT build the tagset.', file=sys.stderr)
else:
print('We WILL build the tagset (for partitioning/traversal).',
file=sys.stderr)
print('making nodegraph', file=sys.stderr)
nodegraph = khmer_args.create_nodegraph(args)
oxfuncs.build_graph(filenames, nodegraph, args.threads,
not args.no_build_tagset)
print('Total number of unique k-mers: {0}'.format(
nodegraph.n_unique_kmers()), file=sys.stderr)
print('saving k-mer nodegraph in', base, file=sys.stderr)
nodegraph.save(base)
if not args.no_build_tagset:
print('saving tagset in', base + '.tagset', file=sys.stderr)
nodegraph.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % nodegraph.n_unique_kmers())
fp_rate = \
khmer.calc_expected_collisions(
nodegraph, args.force, max_false_pos=.15)
# 0.18 is ACTUAL MAX. Do not change.
print('false positive rate estimated to be %1.3f' % fp_rate,
file=sys.stderr)
print('\nfalse positive rate estimated to be %1.3f' % fp_rate,
file=info_fp)
print('wrote to ' + base + '.info and ' + base, file=sys.stderr)
if not args.no_build_tagset:
print('and ' + base + '.tagset', file=sys.stderr)
sys.exit(0)
def main():
parser = build_construct_args()
parser.add_argument('--build-tagset',
'-t',
default=True,
action='store_false',
help='Construct tagset while loading sequences')
parser.add_argument('output_filename')
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print >> sys.stderr, "** WARNING: hashsize is default! You absodefly want to increase this!\n** Please read the docs!"
print >> sys.stderr, '\nPARAMETERS:'
print >> sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >> sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >> sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print >> sys.stderr, ''
print >> sys.stderr, 'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize / 8)' % (
args.n_hashes * args.min_hashsize / 8.)
print >> sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
base = args.output_filename
filenames = args.input_filenames
print 'Saving hashtable to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
###
print 'making hashtable'
ht = khmer.new_hashbits(K, HT_SIZE, N_HT)
for n, filename in enumerate(filenames):
print 'consuming input', filename
ht.consume_fasta_and_tag(filename)
print 'saving hashtable in', base + '.ht'
ht.save(base + '.ht')
print 'saving tagset in', base + '.tagset'
ht.save_tagset(base + '.tagset')
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >> sys.stderr, "**"
print >> sys.stderr, "** ERROR: the graph structure is too small for"
print >> sys.stderr, "** this data set. Increase hashsize/num ht."
print >> sys.stderr, "**"
sys.exit(-1)
def main():
info('optimal_args_hashbits.py', ['graph', 'SeqAn'])
args = get_parser().parse_args()
report_on_config(args, hashtype='hashbits')
filenames = args.input_filenames
base = filenames[0]
for _ in args.input_filenames:
check_input_files(_, False)
check_space(args.input_filenames, False)
print('Counting kmers from sequences in %s' % repr(filenames),
file=sys.stderr)
htable = khmer.new_hashbits(args.ksize, args.max_tablesize, args.n_tables)
target_method = htable.consume_fasta_with_reads_parser
for _, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
print('consuming input', filename, file=sys.stderr)
for num in xrange(args.threads):
cur_thread = threading.Thread(target=target_method,
args=(rparser, ))
threads.append(cur_thread)
cur_thread.start()
for thread in threads:
thread.join()
unique_kmers = htable.n_unique_kmers()
print('Total number of unique k-mers: {0}'.format(unique_kmers),
file=sys.stderr)
info_optimal = open(base + '.optimal_args', 'w')
fp_rate = khmer.calc_expected_collisions(htable)
print('fp rate estimated to be %1.3f' % fp_rate, file=sys.stderr)
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print("**", file=sys.stderr)
print(
"** ERROR: the graph structure is too small for this data set."
"Increase table size/# tables.",
file=sys.stderr)
print("**", file=sys.stderr)
if not False:
sys.exit(1)
to_print = output_gen(unique_kmers, fp_rate)
print(to_print, file=info_optimal)
print('optimal arguments were written to',
base + '.optimal_args',
file=sys.stderr)
def main():
parser = build_construct_args()
add_threading_args(parser)
parser.add_argument('datafile')
args = parser.parse_args()
report_on_config(args)
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
n_threads = int(args.n_threads)
config = khmer.get_config()
bufsz = config.get_reads_input_buffer_size()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT, n_threads)
filename = args.datafile
### first, load reads into hash table
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input, round 1 --', filename
for tnum in xrange(n_threads):
t = \
threading.Thread(
target=ht.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(t)
t.start()
for t in threads:
t.join()
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
### now, count.
total = 0
total_unique = 0
for n, record in enumerate(screed.open(filename)):
total += 1
last_kmer = record.sequence[-K:]
count = ht.get(last_kmer)
if count == 1:
total_unique += 1
print 'singletons: %d unique; of %d total; %.3f' % \
(total_unique, total, total_unique/float(total))
def main():
parser = build_construct_args()
parser.add_argument('--build-tagset', '-t', default=True,
action='store_false',
help='Construct tagset while loading sequences')
parser.add_argument('output_filename')
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print>>sys.stderr, "** WARNING: hashsize is default! You absodefly want to increase this!\n** Please read the docs!"
print>>sys.stderr, '\nPARAMETERS:'
print>>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print>>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print>>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print>>sys.stderr, ''
print>>sys.stderr, 'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize / 8)' % (args.n_hashes * args.min_hashsize / 8.)
print>>sys.stderr, '-'*8
K=args.ksize
HT_SIZE=args.min_hashsize
N_HT=args.n_hashes
base = args.output_filename
filenames = args.input_filenames
print 'Saving hashtable to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
###
print 'making hashtable'
ht = khmer.new_hashbits(K, HT_SIZE, N_HT)
for n, filename in enumerate(filenames):
print 'consuming input', filename
ht.consume_fasta_and_tag(filename)
print 'saving hashtable in', base + '.ht'
ht.save(base + '.ht')
print 'saving tagset in', base + '.tagset'
ht.save_tagset(base + '.tagset')
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the graph structure is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
sys.exit(-1)
def main():
info('optimal_args_nodegraph.py', ['graph', 'SeqAn'])
args = get_parser().parse_args()
report_on_config(args, graphtype='nodegraph')
filenames = args.input_filenames
base = filenames[0]
for _ in args.input_filenames:
check_input_files(_, False)
check_space(args.input_filenames, False)
print('Counting kmers from sequences in %s' % repr(filenames),
file=sys.stderr)
htable = khmer.new_nodegraph(args.ksize, args.max_tablesize, args.n_tables)
target_method = htable.consume_fasta_with_reads_parser
for _, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
print('consuming input', filename, file=sys.stderr)
for num in xrange(args.threads):
cur_thread = threading.Thread(
target=target_method, args=(rparser,))
threads.append(cur_thread)
cur_thread.start()
for thread in threads:
thread.join()
unique_kmers = htable.n_unique_kmers()
print('Total number of unique k-mers: {0}'.format(unique_kmers),
file=sys.stderr)
info_optimal = open(base + '.optimal_args', 'w')
fp_rate = khmer.calc_expected_collisions(htable)
print('fp rate estimated to be %1.3f' % fp_rate, file=sys.stderr)
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print("**", file=sys.stderr)
print("** ERROR: the graph structure is too small for this data set."
"Increase table size/# tables.", file=sys.stderr)
print("**", file=sys.stderr)
if not False:
sys.exit(1)
to_print = graphsize_args_report(unique_kmers, fp_rate)
print(to_print, file=info_optimal)
print('optimal arguments were written to', base + '.optimal_args',
file=sys.stderr)
def main():
parser = build_construct_args()
add_threading_args(parser)
parser.add_argument('datafile')
args = parser.parse_args()
report_on_config(args)
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
n_threads = int(args.n_threads)
config = khmer.get_config()
bufsz = config.get_reads_input_buffer_size()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT, n_threads)
filename = args.datafile
### first, load reads into hash table
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input, round 1 --', filename
for tnum in xrange(n_threads):
t = \
threading.Thread(
target=ht.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(t)
t.start()
for t in threads:
t.join()
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
### now, count.
total = 0
total_unique = 0
for n, record in enumerate(screed.open(filename)):
total += 1
last_kmer = record.sequence[-K:]
count = ht.get(last_kmer)
if count == 1:
total_unique += 1
print 'singletons: %d unique; of %d total; %.3f' % \
(total_unique, total, total_unique/float(total))
def main(args):
info('build-graph.py', ['graph', 'SeqAn'])
report_on_config(args, hashtype='hashbits')
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
check_space(args.input_filenames, args.force)
check_space_for_hashtable(
(float(args.n_tables * args.min_tablesize) / 8.), args.force)
print >>sys.stderr, 'Saving k-mer presence table to %s' % base
print >>sys.stderr, 'Loading kmers from sequences in %s' % repr(filenames)
if args.no_build_tagset:
print >>sys.stderr, 'We WILL NOT build the tagset.'
else:
print >>sys.stderr, 'We WILL build the tagset', \
' (for partitioning/traversal).'
print >>sys.stderr, 'making k-mer presence table'
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
functions.build_graph(filenames, htable, args.threads,
not args.no_build_tagset)
print >> sys.stderr, 'Total number of unique k-mers: {0}'.format(
htable.n_unique_kmers())
print >>sys.stderr, 'saving k-mer presence table in', base + '.pt'
htable.save(base + '.pt')
if not args.no_build_tagset:
print >>sys.stderr, 'saving tagset in', base + '.tagset'
htable.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % htable.n_unique_kmers())
fp_rate = \
khmer.calc_expected_collisions(htable, args.force, max_false_pos=.15)
# 0.18 is ACTUAL MAX. Do not change.
print >>sys.stderr, 'false positive rate estimated to be %1.3f' % fp_rate
print >>info_fp, '\nfalse positive rate estimated to be %1.3f' % fp_rate
print >> sys.stderr, 'wrote to', base + '.info and', base + '.pt'
if not args.no_build_tagset:
print >> sys.stderr, 'and ' + base + '.tagset'
sys.exit(0)
def main(args):
info("build-graph.py", ["graph", "SeqAn"])
report_on_config(args, hashtype="hashbits")
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
check_space(args.input_filenames, args.force)
check_space_for_hashtable((float(args.n_tables * args.min_tablesize) / 8.0), args.force)
print("Saving k-mer presence table to %s" % base, file=sys.stderr)
print("Loading kmers from sequences in %s" % repr(filenames), file=sys.stderr)
if args.no_build_tagset:
print("We WILL NOT build the tagset.", file=sys.stderr)
else:
print("We WILL build the tagset (for partitioning/traversal).", file=sys.stderr)
print("making k-mer presence table", file=sys.stderr)
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
functions.build_graph(filenames, htable, args.threads, not args.no_build_tagset)
print("Total number of unique k-mers: {0}".format(htable.n_unique_kmers()), file=sys.stderr)
print("saving k-mer presence table in", base + ".pt", file=sys.stderr)
htable.save(base + ".pt")
if not args.no_build_tagset:
print("saving tagset in", base + ".tagset", file=sys.stderr)
htable.save_tagset(base + ".tagset")
info_fp = open(base + ".info", "w")
info_fp.write("%d unique k-mers" % htable.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(htable, args.force, max_false_pos=0.15)
# 0.18 is ACTUAL MAX. Do not change.
print("false positive rate estimated to be %1.3f" % fp_rate, file=sys.stderr)
print("\nfalse positive rate estimated to be %1.3f" % fp_rate, file=info_fp)
print("wrote to", base + ".info and", base + ".pt", file=sys.stderr)
if not args.no_build_tagset:
print("and " + base + ".tagset", file=sys.stderr)
sys.exit(0)
def main(args):
info("build-graph.py", ["graph", "SeqAn"])
report_on_config(args, graphtype="nodegraph")
base = args.output_filename
filenames = args.input_filenames
for fname in args.input_filenames:
check_input_files(fname, args.force)
graphsize = calculate_graphsize(args, "nodegraph")
check_space_for_graph(args.output_filename, graphsize, args.force)
print("Saving k-mer nodegraph to %s" % base, file=sys.stderr)
print("Loading kmers from sequences in %s" % repr(filenames), file=sys.stderr)
if args.no_build_tagset:
print("We WILL NOT build the tagset.", file=sys.stderr)
else:
print("We WILL build the tagset (for partitioning/traversal).", file=sys.stderr)
print("making nodegraph", file=sys.stderr)
nodegraph = khmer_args.create_nodegraph(args)
oxfuncs.build_graph(filenames, nodegraph, args.threads, not args.no_build_tagset)
print("Total number of unique k-mers: {0}".format(nodegraph.n_unique_kmers()), file=sys.stderr)
print("saving k-mer nodegraph in", base, file=sys.stderr)
nodegraph.save(base)
if not args.no_build_tagset:
print("saving tagset in", base + ".tagset", file=sys.stderr)
nodegraph.save_tagset(base + ".tagset")
info_fp = open(base + ".info", "w")
info_fp.write("%d unique k-mers" % nodegraph.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(nodegraph, args.force, max_false_pos=0.15)
# 0.18 is ACTUAL MAX. Do not change.
print("false positive rate estimated to be %1.3f" % fp_rate, file=sys.stderr)
print("\nfalse positive rate estimated to be %1.3f" % fp_rate, file=info_fp)
print("wrote to " + base + ".info and " + base, file=sys.stderr)
if not args.no_build_tagset:
print("and " + base + ".tagset", file=sys.stderr)
sys.exit(0)
def main(): # pylint: disable=too-many-branches,too-many-statements
info('saturate-by-median.py', ['diginorm'])
args = get_parser().parse_args()
report_on_config(args)
report_fp = args.report
report_frequency = args.report_frequency
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames)
if args.savetable:
check_space_for_hashtable(args.n_tables * args.min_tablesize)
# list to save error files along with throwing exceptions
if args.force:
corrupt_files = []
if args.loadtable:
print 'loading k-mer counting table from', args.loadtable
htable = khmer.load_counting_hash(args.loadtable)
else:
print 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables)
total = 0
discarded = 0
for index, input_filename in enumerate(args.input_filenames):
total_acc = 0
discarded_acc = 0
try:
total_acc, discarded_acc = normalize_by_median(
input_filename, htable, args, report_fp, report_frequency)
except IOError as err:
handle_error(err, input_filename)
if not args.force:
print >> sys.stderr, '** Exiting!'
sys.exit(1)
else:
print >> sys.stderr, '*** Skipping error file, moving on...'
corrupt_files.append(input_filename)
else:
if total_acc == 0 and discarded_acc == 0:
print 'SKIPPED empty file', input_filename
else:
total += total_acc
discarded += discarded_acc
print 'DONE with {inp}; kept {kept} of {total} or {perc:2}%'\
.format(inp=input_filename,
kept=total - discarded, total=total,
perc=int(100. - discarded / float(total) * 100.))
if args.savetable:
print 'Saving k-mer counting table through', input_filename
print '...saving to', args.savetable
htable.save(args.savetable)
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
if args.force and len(corrupt_files) > 0:
print >> sys.stderr, "** WARNING: Finished with errors!"
print >> sys.stderr, "** IOErrors occurred in the following files:"
print >> sys.stderr, "\t", " ".join(corrupt_files)
if fp_rate > MAX_FALSE_POSITIVE_RATE:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" for this data set. Increase tablesize/# "
"tables.")
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
sys.exit(1)
def main():
info('load-into-counting.py', ['counting'])
args = get_parser().parse_args()
report_on_config(args)
base = args.output_countingtable_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_file_status(name)
check_space(args.input_sequence_filename)
check_space_for_hashtable(args.n_tables * args.min_tablesize)
print 'Saving k-mer counting table to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
print 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables, args.n_threads)
htable.set_use_bigcount(args.bigcount)
config = khmer.get_config()
config.set_reads_input_buffer_size(args.n_threads * 64 * 1024)
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename, args.n_threads)
threads = []
print 'consuming input', filename
for _ in xrange(args.n_threads):
cur_thrd = \
threading.Thread(
target=htable.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for _ in threads:
_.join()
if index > 0 and index % 10 == 0:
check_space_for_hashtable(args.n_tables * args.min_tablesize)
print 'mid-save', base
htable.save(base)
open(base + '.info', 'w').write('through %s' % filename)
print 'saving', base
htable.save(base)
info_fp = open(base + '.info', 'w')
info_fp.write('through end: %s\n' % filename)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
print >> info_fp, 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" this data set. Increase tablesize/# tables.")
print >> sys.stderr, "**"
sys.exit(1)
print 'DONE.'
def main():
parser = build_construct_args()
parser.add_argument('-C',
'--cutoff',
type=int,
dest='cutoff',
default=DEFAULT_DESIRED_COVERAGE)
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash', default='')
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print >> sys.stderr, "** WARNING: hashsize is default! You absodefly want to increase this!\n** Please read the docs!"
print >> sys.stderr, '\nPARAMETERS:'
print >> sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >> sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >> sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print >> sys.stderr, ''
print >> sys.stderr, 'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize)' % (
args.n_hashes * args.min_hashsize)
print >> sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
DESIRED_COVERAGE = args.cutoff
filenames = args.input_filenames
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keepkad'
outfp = open(output_name, 'w')
for n, record in enumerate(screed.open(input_filename)):
if n > 0 and n % 10000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
total += 1
if len(record.sequence) < K:
continue
seq = record.sequence.replace('N', 'A')
kad = ht.get_kadian_count(seq)
if kad < DESIRED_COVERAGE:
ht.consume(seq)
outfp.write('>%s\n%s\n' % (record.name, record.sequence))
else:
discarded += 1
print 'DONE with', input_filename, '; kept', total - discarded, 'of',\
total, 'or', int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >> sys.stderr, "**"
print >> sys.stderr, "** ERROR: the counting hash is too small for"
print >> sys.stderr, "** this data set. Increase hashsize/num ht."
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main(): # pylint: disable=too-many-branches,too-many-statements
parser = sanitize_help(get_parser())
args = parser.parse_args()
configure_logging(args.quiet)
report_on_config(args)
report_fp = args.report
force_single = args.force_single
# check for similar filenames
# if we're using a single output file only check for identical filenames
# otherwise, check for identical BASE names as well.
filenames = []
basenames = []
for pathfilename in args.input_filenames:
filenames.append(pathfilename)
if args.single_output_file:
continue # nothing more to worry about
basename = os.path.basename(pathfilename)
if basename in basenames:
log_error('ERROR: Duplicate filename--Cannot handle this!')
log_error('** Exiting!')
sys.exit(1)
basenames.append(basename)
# check that files exist and there is sufficient output disk space.
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savegraph is not None:
graphsize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.savegraph, graphsize, args.force)
# load or create counting table.
if args.loadgraph:
log_info('loading k-mer countgraph from {graph}', graph=args.loadgraph)
countgraph = khmer.load_countgraph(args.loadgraph)
else:
log_info('making countgraph')
countgraph = khmer_args.create_countgraph(args)
# create an object to handle diginorm of all files
norm = Normalizer(args.cutoff, countgraph)
with_diagnostics = WithDiagnostics(norm, report_fp, args.report_frequency)
# make a list of all filenames and if they're paired or not;
# if we don't know if they're paired, default to allowing but not
# forcing pairing.
files = []
for element in filenames:
files.append([element, args.paired])
if args.unpaired_reads:
files.append([args.unpaired_reads, False])
corrupt_files = []
outfp = None
output_name = None
if args.single_output_file:
outfp = get_file_writer(args.single_output_file, args.gzip, args.bzip)
else:
if '-' in filenames or '/dev/stdin' in filenames:
print(
"Accepting input from stdin; output filename must "
"be provided with '-o'.",
file=sys.stderr)
sys.exit(1)
#
# main loop: iterate over all files given, do diginorm.
#
for filename, require_paired in files:
if not args.single_output_file:
output_name = os.path.basename(filename) + '.keep'
outfp = open(output_name, 'wb')
outfp = get_file_writer(outfp, args.gzip, args.bzip)
# failsafe context manager in case an input file breaks
with catch_io_errors(filename, outfp, args.single_output_file,
args.force, corrupt_files):
screed_iter = clean_input_reads(screed.open(filename))
reader = broken_paired_reader(screed_iter,
min_length=args.ksize,
force_single=force_single,
require_paired=require_paired)
# actually do diginorm
for record in with_diagnostics(reader, filename):
if record is not None:
write_record(record, outfp)
log_info('output in {name}', name=describe_file_handle(outfp))
if not args.single_output_file:
outfp.close()
# finished - print out some diagnostics.
log_info('Total number of unique k-mers: {umers}',
umers=countgraph.n_unique_kmers())
if args.savegraph is not None:
log_info('...saving to {name}', name=args.savegraph)
countgraph.save(args.savegraph)
fp_rate = \
khmer.calc_expected_collisions(countgraph, False, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
log_info('fp rate estimated to be {fpr:1.3f}', fpr=fp_rate)
if args.force and len(corrupt_files) > 0:
log_error("** WARNING: Finished with errors!")
log_error("** I/O Errors occurred in the following files:")
log_error("\t" + " ".join(corrupt_files))
def main():
info('load-into-counting.py', ['counting', 'SeqAn'])
args = sanitize_help(get_parser()).parse_args()
report_on_config(args)
base = args.output_countgraph_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_input_files(name, args.force)
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.output_countgraph_filename, tablesize,
args.force)
check_file_writable(base)
check_file_writable(base + ".info")
print('Saving k-mer countgraph to %s' % base, file=sys.stderr)
print('Loading kmers from sequences in %s' % repr(filenames),
file=sys.stderr)
# clobber the '.info' file now, as we always open in append mode below
if os.path.exists(base + '.info'):
os.remove(base + '.info')
print('making countgraph', file=sys.stderr)
countgraph = khmer_args.create_countgraph(args)
countgraph.set_use_bigcount(args.bigcount)
filename = None
total_num_reads = 0
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
print('consuming input', filename, file=sys.stderr)
for _ in range(args.threads):
cur_thrd = \
threading.Thread(
target=countgraph.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
if index > 0 and index % 10 == 0:
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(base, tablesize, args.force)
print('mid-save', base, file=sys.stderr)
countgraph.save(base)
with open(base + '.info', 'a') as info_fh:
print('through', filename, file=info_fh)
total_num_reads += rparser.num_reads
n_kmers = countgraph.n_unique_kmers()
print('Total number of unique k-mers:', n_kmers, file=sys.stderr)
with open(base + '.info', 'a') as info_fp:
print('Total number of unique k-mers:', n_kmers, file=info_fp)
print('saving', base, file=sys.stderr)
countgraph.save(base)
# Change max_false_pos=0.2 only if you really grok it. HINT: You don't
fp_rate = \
khmer.calc_expected_collisions(
countgraph, args.force, max_false_pos=.2)
with open(base + '.info', 'a') as info_fp:
print('fp rate estimated to be %1.3f\n' % fp_rate, file=info_fp)
if args.summary_info:
mr_fmt = args.summary_info.lower()
mr_file = base + '.info.' + mr_fmt
print("Writing summmary info to", mr_file, file=sys.stderr)
with open(mr_file, 'w') as mr_fh:
if mr_fmt == 'json':
mr_data = {
"ht_name": os.path.basename(base),
"fpr": fp_rate,
"num_kmers": n_kmers,
"files": filenames,
"mrinfo_version": "0.2.0",
"num_reads": total_num_reads,
}
json.dump(mr_data, mr_fh)
mr_fh.write('\n')
elif mr_fmt == 'tsv':
mr_fh.write("ht_name\tfpr\tnum_kmers\tnum_reads\tfiles\n")
vals = [
os.path.basename(base),
"{:1.3f}".format(fp_rate),
str(n_kmers),
str(total_num_reads),
";".join(filenames),
]
mr_fh.write("\t".join(vals) + "\n")
print('fp rate estimated to be %1.3f' % fp_rate, file=sys.stderr)
print('DONE.', file=sys.stderr)
print('wrote to:', base + '.info', file=sys.stderr)
def main():
parser = argparse.ArgumentParser(description='XXX')
env_ksize = os.environ.get('KHMER_KSIZE', DEFAULT_K)
env_n_hashes = os.environ.get('KHMER_N_HASHES', DEFAULT_N_HT)
env_hashsize = os.environ.get('KHMER_MIN_HASHSIZE', DEFAULT_MIN_HASHSIZE)
parser.add_argument('--ksize', '-k', type=int, dest='ksize',
default=env_ksize,
help='k-mer size to use')
parser.add_argument('--n_hashes', '-N', type=int, dest='n_hashes',
default=env_n_hashes,
help='number of hash tables to use')
parser.add_argument('--hashsize', '-x', type=float, dest='min_hashsize',
default=env_hashsize,
help='lower bound on hashsize to use')
parser.add_argument('--cutoff', '-C', type=int, dest='abund_cutoff',
help='remove k-mers below this abundance',
default=DEFAULT_CUTOFF)
parser.add_argument('--normalize-to', '-Z', type=int, dest='normalize_to',
help='base cutoff on median k-mer abundance of this',
default=DEFAULT_NORMALIZE_LIMIT)
parser.add_argument('--variable-coverage', '-V', action='store_true',
dest='variable_coverage', default=False,
help='Only trim low-abundance k-mers from sequences '
'that have high coverage.')
parser.add_argument('--tempdir', '-T', type=str, dest='tempdir',
default='./')
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
CUTOFF = args.abund_cutoff
NORMALIZE_LIMIT = args.normalize_to
print >>sys.stderr, "K:", K
print >>sys.stderr, "HT SIZE:", HT_SIZE
print >>sys.stderr, "N HT:", N_HT
print >>sys.stderr, "CUTOFF:", CUTOFF
print >>sys.stderr, "NORMALIZE_LIMIT:", NORMALIZE_LIMIT
print >>sys.stderr, 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print >>sys.stderr, 'created temporary directory %s; ' % tempdir + \
'use -T to change location'
###
save_pass2 = 0
read_bp = 0
read_reads = 0
pass2list = []
for filename in args.input_filenames:
pass2filename = os.path.basename(filename) + '.pass2'
pass2filename = os.path.join(tempdir, pass2filename)
pass2list.append((filename, pass2filename))
pass2fp = open(pass2filename, 'w')
for n, read in enumerate(screed.open(filename)):
if n % 100000 == 0:
print >>sys.stderr, '...', n, filename, save_pass2, \
read_reads, read_bp
read_reads += 1
read_bp += len(read.sequence)
seq = read.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
# has this portion of the graph saturated? if not,
# consume & save => pass2.
if med < NORMALIZE_LIMIT:
ht.consume(seq)
pass2fp.write(output_single(read))
save_pass2 += 1
else:
posns = ht.find_spectral_error_positions(seq, CUTOFF)
posns = add_n_posns(posns, read.sequence)
print read.name, ",".join(map(str, posns))
pass2fp.close()
print >>sys.stderr, '%s: kept aside %d of %d from first pass, in %s' %\
(filename, save_pass2, n + 1, filename)
n_omitted = 0
for orig_filename, pass2filename in pass2list:
print >>sys.stderr,'second pass: looking at ' + \
'sequences kept aside in %s' % pass2filename
for n, read in enumerate(screed.open(pass2filename)):
if n % 100000 == 0:
print >>sys.stderr, '... x 2', n, pass2filename, read_reads, \
read_bp
seq = read.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
if med >= NORMALIZE_LIMIT or not args.variable_coverage:
posns = ht.find_spectral_error_positions(seq, CUTOFF)
posns = add_n_posns(posns, read.sequence)
print read.name, ",".join(map(str, posns))
if args.variable_coverage and med < NORMALIZE_LIMIT:
print read.name, 'V'
n_omitted += 1
print >>sys.stderr, 'removing %s' % pass2filename
os.unlink(pass2filename)
print >>sys.stderr, 'removing temp directory & contents (%s)' % tempdir
shutil.rmtree(tempdir)
print >>sys.stderr, 'read %d reads, %d bp' % (read_reads, read_bp,)
if args.variable_coverage:
print >>sys.stderr, 'omitted %d reads for -V' % (n_omitted)
fp_rate = khmer.calc_expected_collisions(ht)
print >>sys.stderr, \
'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
if fp_rate > MAX_FALSE_POSITIVE_RATE:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" for this data set. Increase tablesize/# "
"tables.")
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
sys.exit(1)
def main():
parser = build_construct_args()
parser.add_argument('-C', '--cutoff', type=int, dest='cutoff',
default=DEFAULT_DESIRED_COVERAGE)
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash',
default='')
parser.add_argument('-R', '--report-to-file', dest='report_file',
type=argparse.FileType('w'))
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print>>sys.stderr, "** WARNING: hashsize is default! You absodefly want to increase this!\n** Please read the docs!"
print>>sys.stderr, '\nPARAMETERS:'
print>>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print>>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print>>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print>>sys.stderr, ''
print>>sys.stderr, 'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize)' % (args.n_hashes * args.min_hashsize)
print>>sys.stderr, '-'*8
K=args.ksize
HT_SIZE=args.min_hashsize
N_HT=args.n_hashes
DESIRED_COVERAGE=args.cutoff
report_fp = args.report_file
filenames = args.input_filenames
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keep'
outfp = open(output_name, 'w')
n = -1
for n, record in enumerate(screed.open(input_filename)):
if n > 0 and n % 100000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
if report_fp:
print>>report_fp, total, total - discarded, \
1. - (discarded / float(total))
report_fp.flush()
total += 1
if len(record.sequence) < K:
continue
seq = record.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
if med < DESIRED_COVERAGE:
ht.consume(seq)
outfp.write('>%s\n%s\n' % (record.name, record.sequence))
else:
discarded += 1
if -1 < n:
print 'DONE with', input_filename, '; kept', total - discarded, 'of',\
total, 'or', int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
else: print 'SKIPPED empty file', input_filename
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
print >>sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main():
parser = build_construct_args()
parser.add_argument('-C', '--cutoff', type=int, dest='cutoff',
default=DEFAULT_DESIRED_COVERAGE)
parser.add_argument('-p', '--paired', action='store_true')
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash',
default='')
parser.add_argument('-R', '--report-to-file', dest='report_file',
type=argparse.FileType('w'))
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print >>sys.stderr, \
"** WARNING: hashsize is default! " \
"You absodefly want to increase this!\n** " \
"Please read the docs!"
print >>sys.stderr, '\nPARAMETERS:'
print >>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >>sys.stderr, \
' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print >>sys.stderr, ' - paired = %s \t\t(-p)' % args.paired
print >>sys.stderr, ''
print >>sys.stderr, \
'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize)' \
% (args.n_hashes * args.min_hashsize)
print >>sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
DESIRED_COVERAGE = args.cutoff
report_fp = args.report_file
filenames = args.input_filenames
# In paired mode we read two records at a time
batch_size = 1
if args.paired:
batch_size = 2
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keep'
outfp = open(output_name, 'w')
n = -1
for n, batch in enumerate(batchwise(screed.open(
input_filename), batch_size)):
if n > 0 and n % 10000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
if report_fp:
print>>report_fp, total, total - discarded, \
1. - (discarded / float(total))
report_fp.flush()
total += batch_size
# If in paired mode, check that the reads are properly interleaved
if args.paired:
if not validpair(batch[0], batch[1]):
print >>sys.stderr, \
'Error: Improperly interleaved pairs %s %s' \
% (batch[0].name, batch[1].name)
sys.exit(-1)
# Emit the batch of reads if any read passes the filter
# and all reads are longer than K
passed_filter = False
passed_length = True
for record in batch:
if len(record.sequence) < K:
passed_length = False
continue
seq = record.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
if med < DESIRED_COVERAGE:
ht.consume(seq)
passed_filter = True
# Emit records if any passed
if passed_length and passed_filter:
for record in batch:
if hasattr(record, 'accuracy'):
outfp.write('@%s\n%s\n+\n%s\n' % (record.name,
record.sequence,
record.accuracy))
else:
outfp.write(
'>%s\n%s\n' % (record.name, record.sequence))
else:
discarded += batch_size
if -1 < n:
print \
'DONE with', input_filename, '; kept', total - discarded, \
'of', total, 'or', \
int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
else:
print 'SKIPPED empty file', input_filename
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
print >>sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main():
info('load-into-counting.py', ['counting'])
args = get_parser().parse_args()
report_on_config(args)
base = args.output_countingtable_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_file_status(name)
check_space(args.input_sequence_filename)
check_space_for_hashtable(args.n_tables * args.min_tablesize)
print 'Saving k-mer counting table to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
print 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables, args.n_threads)
htable.set_use_bigcount(args.bigcount)
config = khmer.get_config()
config.set_reads_input_buffer_size(args.n_threads * 64 * 1024)
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename, args.n_threads)
threads = []
print 'consuming input', filename
for _ in xrange(args.n_threads):
cur_thrd = \
threading.Thread(
target=htable.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for _ in threads:
_.join()
if index > 0 and index % 10 == 0:
check_space_for_hashtable(args.n_tables * args.min_tablesize)
print 'mid-save', base
htable.save(base)
open(base + '.info', 'w').write('through %s' % filename)
print 'saving', base
htable.save(base)
info_fp = open(base + '.info', 'w')
info_fp.write('through end: %s\n' % filename)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
print >> info_fp, 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" this data set. Increase tablesize/# tables.")
print >> sys.stderr, "**"
sys.exit(1)
print 'DONE.'
def main(): # pylint: disable=too-many-branches,too-many-statements
info('saturate-by-median.py', ['diginorm'])
parser = sanitize_help(get_parser())
args = parser.parse_args()
report_on_config(args)
report_fp = args.report
report_frequency = args.report_frequency
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, False)
if args.savegraph:
check_space_for_graph(args, 'countgraph', False)
# list to save error files along with throwing exceptions
if args.force:
corrupt_files = []
if args.loadgraph:
print('loading k-mer countgraph from', args.loadgraph)
htable = khmer.load_countgraph(args.loadgraph)
else:
print('making countgraph')
htable = create_countgraph(args)
total = 0
discarded = 0
for index, input_filename in enumerate(args.input_filenames):
total_acc = 0
discarded_acc = 0
try:
total_acc, discarded_acc = normalize_by_median(input_filename,
htable, args,
report_fp,
report_frequency)
except IOError as err:
handle_error(err, input_filename)
if not args.force:
print("NOTE: This can be overridden using the --force"
" argument", file=sys.stderr)
print('** Exiting!', file=sys.stderr)
sys.exit(1)
else:
print('*** Skipping error file, moving on...', file=sys.stderr)
corrupt_files.append(input_filename)
else:
if total_acc == 0 and discarded_acc == 0:
print('SKIPPED empty file', input_filename)
else:
total += total_acc
discarded += discarded_acc
print('DONE with {inp}; kept {kept} of {total} or {perc:2}%'\
.format(inp=input_filename,
kept=total - discarded, total=total,
perc=int(100. - discarded / float(total) * 100.)))
if args.savegraph:
print('Saving k-mer countgraph through', input_filename)
print('...saving to', args.savegraph)
htable.save(args.savegraph)
# re: threshold, see Zhang et al.,
# http://arxiv.org/abs/1309.2975
fp_rate = khmer.calc_expected_collisions(htable, args.force, max_false_pos=.8)
print('fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate))
if args.force and len(corrupt_files) > 0:
print("** WARNING: Finished with errors!", file=sys.stderr)
print("** I/O Errors occurred in the following files:", file=sys.stderr)
print("\t", " ".join(corrupt_files), file=sys.stderr)
def main():
parser = argparse.ArgumentParser()
parser.add_argument('seqfiles', nargs='+')
parser.add_argument('-o', '--output', default=None)
parser.add_argument('-k', '--ksize', default=DEFAULT_KSIZE, type=int)
parser.add_argument('-x',
'--tablesize',
default=NODEGRAPH_SIZE,
type=float)
parser.add_argument('--force', action='store_true')
args = parser.parse_args()
assert args.ksize % 2, "ksize must be odd"
assert args.output, "you probably want an output file"
print('building graphs and loading files')
# Create graph, and two stop bloom filters - one for loading, one for
# traversing. Create them all here so that we can error out quickly
# if memory is a problem.
graph = khmer.Nodegraph(args.ksize, args.tablesize, 2)
print(graph.ksize(), graph.hashsizes())
stop_bf = khmer.Nodegraph(args.ksize, args.tablesize, 2)
stop_bf2 = khmer.Nodegraph(args.ksize, args.tablesize, 2)
n = 0
# load in all of the input sequences, one file at a time.
for seqfile in args.seqfiles:
for record in screed.open(seqfile):
n += 1
if n % 10000 == 0:
print('...', seqfile, n)
graph.consume(record.sequence)
# complain if too small set of graphs was used.
fp_rate = khmer.calc_expected_collisions(graph,
args.force,
max_false_pos=.05)
# initialize the object that will track information for us.
pathy = Pathfinder(args.ksize)
print('finding high degree nodes')
degree_nodes = khmer.HashSet(args.ksize)
n = 0
for seqfile in args.seqfiles:
for record in screed.open(seqfile):
n += 1
if n % 10000 == 0:
print('...2', seqfile, n)
# walk across sequences, find all high degree nodes,
# name them and cherish them. Don't do this on identical sequences.
if min(stop_bf2.get_kmer_counts(record.sequence)) == 0:
stop_bf2.consume(record.sequence)
degree_nodes += graph.find_high_degree_nodes(record.sequence)
del stop_bf2
if not len(degree_nodes):
print('no high degree nodes; exiting.')
sys.exit(0)
# get all of the degree > 2 nodes and give them IDs.
for node in degree_nodes:
pathy.new_segment(node)
print('traversing linear segments from', len(degree_nodes), 'nodes')
# now traverse from each high degree nodes into all neighboring nodes,
# seeking adjacencies. if neighbor is high degree node, add it to
# adjacencies; if neighbor is not, then traverse the linear path. also
# track minhashes while we're at it.
for n, k in enumerate(degree_nodes):
if n % 10000 == 0:
print('...', n, 'of', len(degree_nodes))
# retrieve the segment ID of the primary node.
k_id = pathy.segments_r[k]
# find all the neighbors of this high-degree node.
nbh = graph.neighbors(k)
for nk in nbh:
# neighbor is high degree? fine, mark its adjacencies.
if nk in degree_nodes:
nk_id = pathy.segments_r[nk]
pathy.add_adjacency(k_id, nk_id)
else:
# linear! walk it.
traverse_and_mark_linear_paths(graph, nk, stop_bf, pathy,
degree_nodes)
print(len(pathy.segments), 'segments, containing',
sum(pathy.segments.values()), 'nodes')
# save to GML
if args.output:
print('saving to', args.output)
fp = open(args.output, 'w')
w = GmlWriter(fp, [], [])
for k, v in pathy.segments.items():
w.add_vertex(k, v, [])
for k, v in pathy.adjacencies.items():
for edge in v:
w.add_edge(k, edge, [])
def main():
parser = sanitize_help(get_parser())
args = parser.parse_args()
if not args.quiet:
info('trim-low-abund.py', ['streaming'])
configure_logging(args.quiet)
###
if len(set(args.input_filenames)) != len(args.input_filenames):
log_error("Error: Cannot input the same filename multiple times.")
sys.exit(1)
if args.trim_at_coverage != DEFAULT_TRIM_AT_COVERAGE and \
not args.variable_coverage:
log_error("Error: --trim-at-coverage/-Z given, but "
"--variable-coverage/-V not specified.")
sys.exit(1)
if args.diginorm_coverage != DEFAULT_DIGINORM_COVERAGE and \
not args.diginorm:
log_error("Error: --diginorm-coverage given, but "
"--diginorm not specified.")
sys.exit(1)
if args.diginorm and args.single_pass:
log_error("Error: --diginorm and --single-pass are incompatible!\n"
"You probably want to use normalize-by-median.py instead.")
sys.exit(1)
###
report_on_config(args)
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savegraph:
graphsize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.savegraph, graphsize, args.force)
if ('-' in args.input_filenames or '/dev/stdin' in args.input_filenames) \
and not args.output:
log_error("Accepting input from stdin; output filename must "
"be provided with -o.")
sys.exit(1)
if args.loadgraph:
log_info('loading countgraph from {graph}', graph=args.loadgraph)
ct = khmer.load_countgraph(args.loadgraph)
else:
log_info('making countgraph')
ct = khmer_args.create_countgraph(args)
K = ct.ksize()
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
log_info(
'created temporary directory {temp};\n'
'use -T to change location',
temp=tempdir)
trimmer = Trimmer(ct, not args.variable_coverage, args.cutoff,
args.trim_at_coverage)
if args.diginorm:
trimmer.set_diginorm(args.diginorm_coverage)
# ### FIRST PASS ###
save_pass2_total = 0
written_bp = 0
written_reads = 0
# only create the file writer once if outfp is specified; otherwise,
# create it for each file.
if args.output:
trimfp = get_file_writer(args.output, args.gzip, args.bzip)
pass2list = []
for filename in args.input_filenames:
# figure out temporary filename for 2nd pass
pass2filename = os.path.basename(filename) + '.pass2'
pass2filename = os.path.join(tempdir, pass2filename)
pass2fp = open(pass2filename, 'w')
# construct output filenames
if args.output is None:
# note: this will be saved in trimfp.
outfp = open(os.path.basename(filename) + '.abundtrim', 'wb')
# get file handle w/gzip, bzip
trimfp = get_file_writer(outfp, args.gzip, args.bzip)
# record all this info
pass2list.append((filename, pass2filename, trimfp))
# input file stuff: get a broken_paired reader.
screed_iter = screed.open(filename)
paired_iter = broken_paired_reader(screed_iter,
min_length=K,
force_single=args.ignore_pairs)
# main loop through the file.
n_start = trimmer.n_reads
save_start = trimmer.n_saved
watermark = REPORT_EVERY_N_READS
for read in trimmer.pass1(paired_iter, pass2fp):
if (trimmer.n_reads - n_start) > watermark:
log_info(
"... {filename} {n_saved} {n_reads} {n_bp} "
"{w_reads} {w_bp}",
filename=filename,
n_saved=trimmer.n_saved,
n_reads=trimmer.n_reads,
n_bp=trimmer.n_bp,
w_reads=written_reads,
w_bp=written_bp)
watermark += REPORT_EVERY_N_READS
# write out the trimmed/etc sequences that AREN'T going to be
# revisited in a 2nd pass.
write_record(read, trimfp)
written_bp += len(read)
written_reads += 1
pass2fp.close()
log_info("{filename}: kept aside {kept} of {total} from first pass",
filename=filename,
kept=trimmer.n_saved - save_start,
total=trimmer.n_reads - n_start)
# first pass goes across all the data, so record relevant stats...
n_reads = trimmer.n_reads
n_bp = trimmer.n_bp
n_skipped = trimmer.n_skipped
bp_skipped = trimmer.bp_skipped
save_pass2_total = trimmer.n_saved
# ### SECOND PASS. ###
# nothing should have been skipped yet!
assert trimmer.n_skipped == 0
assert trimmer.bp_skipped == 0
if args.single_pass:
pass2list = []
# go back through all the files again.
for _, pass2filename, trimfp in pass2list:
log_info('second pass: looking at sequences kept aside in {pass2}',
pass2=pass2filename)
# note that for this second pass, we don't care about paired
# reads - they will be output in the same order they're read in,
# so pairs will stay together if not orphaned. This is in contrast
# to the first loop. Hence, force_single=True below.
screed_iter = screed.open(pass2filename, parse_description=False)
paired_iter = broken_paired_reader(screed_iter,
min_length=K,
force_single=True)
watermark = REPORT_EVERY_N_READS
for read in trimmer.pass2(paired_iter):
if (trimmer.n_reads - n_start) > watermark:
log_info('... x 2 {a} {b} {c} {d} {e} {f} {g}',
a=trimmer.n_reads - n_start,
b=pass2filename,
c=trimmer.n_saved,
d=trimmer.n_reads,
e=trimmer.n_bp,
f=written_reads,
g=written_bp)
watermark += REPORT_EVERY_N_READS
write_record(read, trimfp)
written_reads += 1
written_bp += len(read)
log_info('removing {pass2}', pass2=pass2filename)
os.unlink(pass2filename)
# if we created our own trimfps, close 'em.
if not args.output:
trimfp.close()
log_info('removing temp directory & contents ({temp})', temp=tempdir)
shutil.rmtree(tempdir)
trimmed_reads = trimmer.trimmed_reads
n_passes = 1.0 + (float(save_pass2_total) / n_reads)
percent_reads_trimmed = float(trimmed_reads + (n_reads - written_reads)) /\
n_reads * 100.0
log_info('read {read} reads, {bp} bp', read=n_reads, bp=n_bp)
log_info('wrote {wr} reads, {wbp} bp', wr=written_reads, wbp=written_bp)
log_info('looked at {st} reads twice ({np:.2f} passes)',
st=save_pass2_total,
np=n_passes)
log_info('removed {r} reads and trimmed {t} reads ({p:.2f}%)',
r=n_reads - written_reads,
t=trimmed_reads,
p=percent_reads_trimmed)
log_info('trimmed or removed {p:.2f}%% of bases ({bp} total)',
p=(1 - (written_bp / float(n_bp))) * 100.0,
bp=n_bp - written_bp)
if args.variable_coverage:
percent_reads_hicov = 100.0 * float(n_reads - n_skipped) / n_reads
log_info('{n} reads were high coverage ({p:.2f}%);',
n=n_reads - n_skipped,
p=percent_reads_hicov)
log_info('skipped {r} reads/{bp} bases because of low coverage',
r=n_skipped,
bp=bp_skipped)
fp_rate = \
khmer.calc_expected_collisions(ct, args.force, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
log_info('fp rate estimated to be {fpr:1.3f}', fpr=fp_rate)
log_info('output in *.abundtrim')
if args.savegraph:
log_info("Saving k-mer countgraph to {graph}", graph=args.savegraph)
ct.save(args.savegraph)
def main():
parser = build_counting_args()
parser.add_argument("-t",
"--trusted-cutoff",
dest="trusted_cutoff",
type=int,
default=3)
parser.add_argument("--bits-theta",
help="Tuning parameter controlling"
"trade off of speed vs alignment sensitivity",
default=1.0,
type=float,
dest="bits_theta")
parser.add_argument('--normalize-to',
'-Z',
type=int,
dest='normalize_to',
help='base cutoff on abundance',
default=DEFAULT_NORMALIZE_LIMIT)
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash', default='')
parser.add_argument('--details-out', dest="details_out")
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
print('\nPARAMETERS:', file=sys.stderr)
print(' - kmer size = %d \t\t(-k)' % args.ksize, file=sys.stderr)
print(' - n hashes = %d \t\t(-N)' % args.n_tables, file=sys.stderr)
print(' - min hashsize = %-5.2g \t(-x)' % \
args.max_tablesize, file=sys.stderr)
print('', file=sys.stderr)
print('Estimated memory usage is %.2g bytes ' \
'(n_hashes x min_hashsize)' % \
(args.n_tables * args.max_tablesize), file=sys.stderr)
print('-' * 8, file=sys.stderr)
K = args.ksize
HT_SIZE = args.max_tablesize
N_HT = args.n_tables
filenames = args.input_filenames
if args.loadhash:
print('loading hashtable from', args.loadhash)
ht = khmer.load_countgraph(args.loadhash)
else:
print('making hashtable')
ht = khmer.Countgraph(K, HT_SIZE, N_HT)
aligner = khmer.ReadAligner(ht, args.trusted_cutoff, args.bits_theta)
if args.details_out is not None:
details_out = open(args.details_out, "w")
else:
details_out = None
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keepvar'
outfp = open(output_name, 'w')
for n, record in enumerate(screed.open(input_filename)):
if n > 0 and n % 10000 == 0:
print('... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%')
print('... in file', input_filename)
total += 1
if len(record.sequence) < K:
continue
seq = record.sequence.upper().replace('N', 'A')
##
# build the alignment...
score, graph_alignment, read_alignment, truncated = \
aligner.align(record.sequence)
# next, decide whether or to keep it.
keep = False
if truncated:
keep = True # keep all truncated alignments - why?
else:
# build a better sequence -- this is the corrected one.
graph_seq = graph_alignment.replace("-", "")
# OR?
#graph_seq = ""
#for i in range(len(graph_alignment)):
# if graph_alignment[i] == "-":
# graph_seq += read_alignment[i]
# else:
# graph_seq += graph_alignment[i]
# get the minimum count for this new sequence
mincount = ht.get_min_count(graph_seq)
if mincount < args.normalize_to:
keep = True
if details_out is not None:
details_out.write(
"+{7}\t{0:0.2f}\t{3}\t{4}\nread: "
"{6}\ngraph_aln: {1}\nread_aln: {2}\nstored_seq:{5}\n"
"".format(score, graph_alignment, read_alignment,
truncated, keep, seq, record.sequence,
record.name))
if keep:
ht.consume(seq)
outfp.write('>%s\n%s\n' % (record.name, record.sequence))
else:
discarded += 1
if total:
print('DONE with', input_filename, \
'; kept', total - discarded, 'of', total, 'or', \
int(100. - discarded / float(total) * 100.), '%')
print('output in', output_name)
if args.savehash:
print('Saving hashfile through', input_filename)
print('...saving to', args.savehash)
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht, args.force, max_false_pos=.2)
print('fp rate estimated to be %1.3f' % fp_rate)
def main(): # pylint: disable=too-many-branches,too-many-statements
info('normalize-by-median.py', ['diginorm'])
args = get_parser().parse_args()
report_on_config(args)
report_fp = args.report
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savetable:
check_space_for_hashtable(
args.n_tables * args.min_tablesize, args.force)
# list to save error files along with throwing exceptions
if args.force:
corrupt_files = []
if args.loadtable:
print 'loading k-mer counting table from', args.loadtable
htable = khmer.load_counting_hash(args.loadtable)
else:
print 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables)
total = 0
discarded = 0
input_filename = None
for index, input_filename in enumerate(args.input_filenames):
if args.single_output_filename != '':
output_name = args.single_output_filename
outfp = open(args.single_output_filename, 'a')
else:
output_name = os.path.basename(input_filename) + '.keep'
outfp = open(output_name, 'w')
total_acc = 0
discarded_acc = 0
try:
total_acc, discarded_acc = normalize_by_median(input_filename,
outfp, htable, args,
report_fp)
except IOError as err:
handle_error(err, output_name, input_filename, args.fail_save,
htable)
if not args.force:
print >> sys.stderr, '** Exiting!'
sys.exit(1)
else:
print >> sys.stderr, '*** Skipping error file, moving on...'
corrupt_files.append(input_filename)
else:
if total_acc == 0 and discarded_acc == 0:
print 'SKIPPED empty file', input_filename
else:
total += total_acc
discarded += discarded_acc
print 'DONE with {inp}; kept {kept} of {total} or {perc:2}%'\
.format(inp=input_filename, kept=total - discarded,
total=total, perc=int(100. - discarded /
float(total) * 100.))
print 'output in', output_name
if (args.dump_frequency > 0 and
index > 0 and index % args.dump_frequency == 0):
print 'Backup: Saving k-mer counting file through', input_filename
if args.savetable:
hashname = args.savetable
print '...saving to', hashname
else:
hashname = 'backup.ct'
print 'Nothing given for savetable, saving to', hashname
htable.save(hashname)
if args.report_total_kmers:
print >> sys.stderr, 'Total number of unique k-mers: {0}'.format(
htable.n_unique_kmers())
if args.savetable:
print 'Saving k-mer counting table through', input_filename
print '...saving to', args.savetable
htable.save(args.savetable)
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
if args.force and len(corrupt_files) > 0:
print >> sys.stderr, "** WARNING: Finished with errors!"
print >> sys.stderr, "** IOErrors occurred in the following files:"
print >> sys.stderr, "\t", " ".join(corrupt_files)
if fp_rate > MAX_FALSE_POSITIVE_RATE:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" for this data set. Increase tablesize/# "
"tables.")
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
if not args.force:
sys.exit(1)
def main():
parser = argparse.ArgumentParser(description='XXX')
env_ksize = os.environ.get('KHMER_KSIZE', DEFAULT_K)
env_n_hashes = os.environ.get('KHMER_N_HASHES', DEFAULT_N_HT)
env_hashsize = os.environ.get('KHMER_MIN_HASHSIZE', DEFAULT_MIN_HASHSIZE)
parser.add_argument('--ksize',
'-k',
type=int,
dest='ksize',
default=env_ksize,
help='k-mer size to use')
parser.add_argument('--n_hashes',
'-N',
type=int,
dest='n_hashes',
default=env_n_hashes,
help='number of hash tables to use')
parser.add_argument('--hashsize',
'-x',
type=float,
dest='min_hashsize',
default=env_hashsize,
help='lower bound on hashsize to use')
parser.add_argument('--cutoff',
'-C',
type=int,
dest='abund_cutoff',
help='remove k-mers below this abundance',
default=DEFAULT_CUTOFF)
parser.add_argument('--normalize-to',
'-Z',
type=int,
dest='normalize_to',
help='base cutoff on median k-mer abundance of this',
default=DEFAULT_NORMALIZE_LIMIT)
parser.add_argument('--variable-coverage',
'-V',
action='store_true',
dest='variable_coverage',
default=False,
help='Only trim low-abundance k-mers from sequences '
'that have high coverage.')
parser.add_argument('--tempdir',
'-T',
type=str,
dest='tempdir',
default='./')
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
###
if len(set(args.input_filenames)) != len(args.input_filenames):
print >>sys.stderr, \
"Error: Cannot input the same filename multiple times."
sys.exit(1)
###
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
CUTOFF = args.abund_cutoff
NORMALIZE_LIMIT = args.normalize_to
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print 'created temporary directory %s; use -T to change location' % tempdir
###
save_pass2_total = 0
read_bp = 0
read_reads = 0
wrote_bp = 0
wrote_reads = 0
trimmed_reads = 0
pass2list = []
for filename in args.input_filenames:
pass2filename = os.path.basename(filename) + '.pass2'
pass2filename = os.path.join(tempdir, pass2filename)
trimfilename = os.path.basename(filename) + '.abundtrim'
pass2list.append((filename, pass2filename, trimfilename))
pass2fp = open(pass2filename, 'w')
trimfp = open(trimfilename, 'w')
save_pass2 = 0
for n, read in enumerate(screed.open(filename)):
if n % 10000 == 0:
print '...', n, filename, save_pass2, read_reads, read_bp, \
wrote_reads, wrote_bp
read_reads += 1
read_bp += len(read.sequence)
seq = read.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
# has this portion of the graph saturated? if not,
# consume & save => pass2.
if med < NORMALIZE_LIMIT:
ht.consume(seq)
pass2fp.write(output_single(read))
save_pass2 += 1
else: # trim!!
trim_seq, trim_at = ht.trim_on_abundance(seq, CUTOFF)
if trim_at >= K:
trimfp.write(output_single(read, trim_at))
wrote_reads += 1
wrote_bp += trim_at
if trim_at != len(read.sequence):
trimmed_reads += 1
pass2fp.close()
trimfp.close()
print '%s: kept aside %d of %d from first pass, in %s' % \
(filename, save_pass2, n, filename)
save_pass2_total += save_pass2
skipped_n = 0
skipped_bp = 0
for orig_filename, pass2filename, trimfilename in pass2list:
print 'second pass: looking at sequences kept aside in %s' % \
pass2filename
for n, read in enumerate(screed.open(pass2filename)):
if n % 10000 == 0:
print '... x 2', n, pass2filename, read_reads, read_bp, \
wrote_reads, wrote_bp
trimfp = open(trimfilename, 'a')
seq = read.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
# do we retain low-abundance components unchanged?
if med < NORMALIZE_LIMIT and args.variable_coverage:
trimfp.write(output_single(read))
wrote_reads += 1
wrote_bp += len(read.sequence)
skipped_n += 1
skipped_bp += len(read.sequence)
# otherwise, examine/trim/truncate.
else: # med >= NORMALIZE LIMIT or not args.variable_coverage
trim_seq, trim_at = ht.trim_on_abundance(seq, CUTOFF)
if trim_at >= K:
trimfp.write(output_single(read, trim_at))
wrote_reads += 1
wrote_bp += trim_at
if trim_at != len(read.sequence):
trimmed_reads += 1
print 'removing %s' % pass2filename
os.unlink(pass2filename)
print 'removing temp directory & contents (%s)' % tempdir
shutil.rmtree(tempdir)
print 'read %d reads, %d bp' % (
read_reads,
read_bp,
)
print 'wrote %d reads, %d bp' % (
wrote_reads,
wrote_bp,
)
print 'removed %d reads and trimmed %d reads' % (
read_reads - wrote_reads,
trimmed_reads,
)
print 'looked at %d reads twice' % (save_pass2_total, )
print 'trimmed or removed %.2f%% of bases (%d total)' % \
((1 - (wrote_bp / float(read_bp))) * 100., read_bp - wrote_bp)
if args.variable_coverage:
print 'skipped %d reads/%d bases because of low coverage' % \
(skipped_n, skipped_bp)
print 'output in *.abundtrim'
fp_rate = khmer.calc_expected_collisions(ht)
print >>sys.stderr, \
'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
if fp_rate > MAX_FALSE_POSITIVE_RATE:
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the k-mer counting table is too small"
" for this data set. Increase tablesize/# "
"tables.")
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
sys.exit(1)
def main(): # pylint: disable=too-many-locals,too-many-statements
info('do-partition.py', ['graph'])
args = get_parser().parse_args()
report_on_config(args, hashtype='hashbits')
for infile in args.input_filenames:
check_file_status(infile)
check_space(args.input_filenames)
print 'Saving k-mer presence table to %s' % args.graphbase
print 'Loading kmers from sequences in %s' % repr(args.input_filenames)
print '--'
print 'SUBSET SIZE', args.subset_size
print 'N THREADS', args.n_threads
print '--'
# load-graph
print 'making k-mer presence table'
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
for _, filename in enumerate(args.input_filenames):
print 'consuming input', filename
htable.consume_fasta_and_tag(filename)
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the graph structure is too small for"
" this data set. Increase k-mer presence table "
"size/num of tables.")
print >> sys.stderr, "**"
sys.exit(1)
# partition-graph
# do we want to exhaustively traverse the graph?
stop_big_traversals = args.no_big_traverse
if stop_big_traversals:
print '** This script brakes for lumps: stop_big_traversals is true.'
else:
print '** Traverse all the things: stop_big_traversals is false.'
#
# now, partition!
#
# divide the tags up into subsets
divvy = htable.divide_tags_into_subsets(int(args.subset_size))
n_subsets = len(divvy)
divvy.append(0)
# build a queue of tasks:
worker_q = Queue.Queue()
# break up the subsets into a list of worker tasks
for _ in range(0, n_subsets):
start = divvy[_]
end = divvy[_ + 1]
worker_q.put((htable, _, start, end))
print 'enqueued %d subset tasks' % n_subsets
open('%s.info' % args.graphbase,
'w').write('%d subsets total\n' % (n_subsets))
if n_subsets < args.n_threads:
args.n_threads = n_subsets
# start threads!
print 'starting %d threads' % args.n_threads
print '---'
threads = []
for _ in range(args.n_threads):
cur_thread = threading.Thread(target=worker,
args=(worker_q, args.graphbase,
stop_big_traversals))
threads.append(cur_thread)
cur_thread.start()
print 'done starting threads'
# wait for threads
for _ in threads:
_.join()
print '---'
print 'done making subsets! see %s.subset.*.pmap' % (args.graphbase, )
# merge-partitions
pmap_files = glob.glob(args.graphbase + '.subset.*.pmap')
print 'loading %d pmap files (first one: %s)' % (len(pmap_files),
pmap_files[0])
htable = khmer.new_hashbits(args.ksize, 1, 1)
for pmap_file in pmap_files:
print 'merging', pmap_file
htable.merge_subset_from_disk(pmap_file)
if args.remove_subsets:
print 'removing pmap files'
for pmap_file in pmap_files:
os.unlink(pmap_file)
# annotate-partitions
for infile in args.input_filenames:
print 'outputting partitions for', infile
outfile = os.path.basename(infile) + '.part'
part_count = htable.output_partitions(infile, outfile)
print 'output %d partitions for %s' % (part_count, infile)
print 'partitions are in', outfile
def main():
parser = build_construct_args()
add_threading_args(parser)
parser.add_argument('output_filename')
parser.add_argument('input_filenames', nargs='+')
parser.add_argument('-b', '--no-bigcount', dest='bigcount', default=True,
action='store_false',
help='Do not count k-mers past 255')
args = parser.parse_args()
report_on_config(args)
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
base = args.output_filename
filenames = args.input_filenames
n_threads = int(args.n_threads)
print 'Saving hashtable to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
###
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT, n_threads)
ht.set_use_bigcount(args.bigcount)
config = khmer.get_config()
bufsz = config.get_reads_input_buffer_size()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
for n, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input', filename
for tnum in xrange(n_threads):
t = \
threading.Thread(
target=ht.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(t)
t.start()
for t in threads:
t.join()
if n > 0 and n % 10 == 0:
print 'mid-save', base
ht.save(base)
open(base + '.info', 'w').write('through %s' % filename)
print 'saving', base
ht.save(base)
info_fp = open(base + '.info', 'w')
info_fp.write('through end: %s\n' % filename)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
print >>info_fp, 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
sys.exit(-1)
print 'DONE.'
def main():
info('collect-reads.py', ['counting'])
args = get_parser().parse_args()
report_on_config(args)
base = args.output_countingtable_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_input_files(name, False)
check_space(args.input_sequence_filename, False)
check_space_for_hashtable(args.n_tables * args.min_tablesize, False)
print 'Saving k-mer counting table to %s' % base
print 'Loading sequences from %s' % repr(filenames)
if args.output:
print 'Outputting sequences to', args.output
print 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize)
htable.set_use_bigcount(args.bigcount)
total_coverage = 0.
n = 0
for index, filename in enumerate(filenames):
for record in screed.open(filename):
seq = record.sequence.upper()
if 'N' in seq:
seq = seq.replace('N', 'A')
try:
med, _, _ = htable.get_median_count(seq)
except ValueError:
continue
total_coverage += med
n += 1
if total_coverage / float(n) > args.coverage:
print 'reached target average coverage:', \
total_coverage / float(n)
break
htable.consume(seq)
if args.output:
args.output.write(output_single(record))
if n % 100000 == 0:
print '...', index, filename, n, total_coverage / float(n)
if total_coverage / float(n) > args.coverage:
break
print 'Collected %d reads' % (n, )
if args.report_total_kmers:
print >> sys.stderr, 'Total number of k-mers: {0}'.format(
htable.n_occupied())
print 'saving', base
htable.save(base)
info_fp = open(base + '.info', 'w')
info_fp.write('through end: %s\n' % filenames[-1])
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(htable,
args.force,
max_false_pos=.2)
print 'fp rate estimated to be %1.3f' % fp_rate
print >> info_fp, 'fp rate estimated to be %1.3f' % fp_rate
print 'DONE.'
def main(): # pylint: disable=too-many-locals,too-many-statements
args = sanitize_help(get_parser()).parse_args()
report_on_config(args, graphtype='nodegraph')
for infile in args.input_filenames:
check_input_files(infile, args.force)
check_space(args.input_filenames, args.force)
print('Saving k-mer nodegraph to %s' % args.graphbase, file=sys.stderr)
print('Loading kmers from sequences in %s' % repr(args.input_filenames),
file=sys.stderr)
print('--', file=sys.stderr)
print('SUBSET SIZE', args.subset_size, file=sys.stderr)
print('N THREADS', args.threads, file=sys.stderr)
print('--', file=sys.stderr)
# load-graph.py
print('making nodegraph', file=sys.stderr)
nodegraph = khmer_args.create_nodegraph(args)
for _, filename in enumerate(args.input_filenames):
print('consuming input', filename, file=sys.stderr)
nodegraph.consume_seqfile_and_tag(filename)
# 0.18 is ACTUAL MAX. Do not change.
fp_rate = \
khmer.calc_expected_collisions(
nodegraph, args.force, max_false_pos=.15)
print('fp rate estimated to be %1.3f' % fp_rate, file=sys.stderr)
# partition-graph
# do we want to exhaustively traverse the graph?
stop_big_traversals = args.no_big_traverse
if stop_big_traversals:
print('** This script brakes for lumps: ',
'stop_big_traversals is true.',
file=sys.stderr)
else:
print('** Traverse all the things:',
' stop_big_traversals is false.',
file=sys.stderr)
#
# now, partition!
#
# divide the tags up into subsets
divvy = nodegraph.divide_tags_into_subsets(int(args.subset_size))
divvy = list(divvy)
n_subsets = len(divvy)
divvy.append(0)
# build a queue of tasks:
worker_q = queue.Queue()
# break up the subsets into a list of worker tasks
for _ in range(0, n_subsets):
start = divvy[_]
end = divvy[_ + 1]
worker_q.put((nodegraph, _, start, end))
print('enqueued %d subset tasks' % n_subsets, file=sys.stderr)
open('%s.info' % args.graphbase,
'w').write('%d subsets total\n' % (n_subsets))
if n_subsets < args.threads:
args.threads = n_subsets
# start threads!
print('starting %d threads' % args.threads, file=sys.stderr)
print('---', file=sys.stderr)
threads = []
for _ in range(args.threads):
cur_thread = threading.Thread(target=worker,
args=(worker_q, args.graphbase,
stop_big_traversals))
threads.append(cur_thread)
cur_thread.start()
print('done starting threads', file=sys.stderr)
# wait for threads
for _ in threads:
_.join()
print('---', file=sys.stderr)
print('done making subsets! see %s.subset.*.pmap' % (args.graphbase, ),
file=sys.stderr)
# merge-partitions
pmap_files = glob.glob(args.graphbase + '.subset.*.pmap')
print('loading %d pmap files (first one: %s)' %
(len(pmap_files), pmap_files[0]),
file=sys.stderr)
nodegraph = khmer.Nodegraph(args.ksize, 1, 1)
for pmap_file in pmap_files:
print('merging', pmap_file, file=sys.stderr)
nodegraph.merge_subset_from_disk(pmap_file)
if not args.keep_subsets:
print('removing pmap files', file=sys.stderr)
for pmap_file in pmap_files:
os.unlink(pmap_file)
# annotate-partitions
for infile in args.input_filenames:
print('outputting partitions for', infile, file=sys.stderr)
outfile = os.path.basename(infile) + '.part'
part_count = nodegraph.output_partitions(infile, outfile)
print('output %d partitions for %s' % (part_count, infile),
file=sys.stderr)
print('partitions are in', outfile, file=sys.stderr)
def main():
parser = build_counting_args()
parser.add_argument("-t",
"--trusted-cutoff",
dest="trusted_cutoff",
type=int,
default=3)
parser.add_argument(
"--bits-theta",
help=
"Tuning parameter controlling trade off of speed vs alignment sensitivity",
default=1.0,
type=float,
dest="bits_theta")
parser.add_argument('-C',
'--cutoff',
type=int,
dest='cutoff',
default=DEFAULT_MINIMUM_COVERAGE)
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash', default='')
parser.add_argument('--details-out', dest="details_out")
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
print >> sys.stderr, '\nPARAMETERS:'
print >> sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >> sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % \
args.min_hashsize
print >> sys.stderr, ''
print >>sys.stderr, 'Estimated memory usage is %.2g bytes ' \
'(n_hashes x min_hashsize)' % (
args.n_hashes * args.min_hashsize)
print >> sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
DESIRED_COVERAGE = args.cutoff
filenames = args.input_filenames
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
aligner = khmer.new_readaligner(ht, args.trusted_cutoff, args.bits_theta)
if args.details_out != None:
details_out = open(args.details_out, "w")
else:
details_out = None
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keepalign'
outfp = open(output_name, 'w')
for n, record in enumerate(screed.open(input_filename)):
if n > 0 and n % 10000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
total += 1
if len(record.sequence) < K:
continue
seq = record.sequence.upper().replace('N', 'A')
##
score, graph_alignment, read_alignment, truncated = aligner.align(
record.sequence)
keep = False
if truncated:
keep = True
else:
if False:
graph_seq = graph_alignment.replace("-", "")
else:
graph_seq = ""
for i in range(len(graph_alignment)):
if graph_alignment[i] == "-":
graph_seq += read_alignment[i]
else:
graph_seq += graph_alignment[i]
mincount = ht.get_min_count(graph_seq)
keep = True
seq = graph_seq
#if mincount < DESIRED_COVERAGE:
# keep = True
# seq = graph_seq
#else:
# assert not keep
if details_out != None:
details_out.write(
"+{7}\t{0:0.2f}\t{3}\t{4}\nread: {6}\ngraph_aln: {1}\nread_aln: {2}\nstored_seq:{5}\n"
.format(score, graph_alignment, read_alignment, truncated,
keep, seq, record.sequence, record.name))
if keep:
ht.consume(seq)
outfp.write('>%s\n%s\n' % (record.name, seq))
else:
discarded += 1
print 'DONE with', input_filename, '; kept', total - discarded, 'of',\
total, 'or', int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >> sys.stderr, "**"
print >> sys.stderr, "** ERROR: the counting hash is too small for"
print >> sys.stderr, "** this data set. Increase hashsize/num ht."
print >> sys.stderr, "**"
print >> sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main():
parser = build_construct_args()
parser.add_argument('-C', '--cutoff', type=int, dest='cutoff',
default=DEFAULT_DESIRED_COVERAGE)
parser.add_argument('-p', '--paired', action='store_true')
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash',
default='')
parser.add_argument('-R', '--report-to-file', dest='report_file',
type=argparse.FileType('w'))
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print>>sys.stderr, "** WARNING: hashsize is default! You absodefly want to increase this!\n** Please read the docs!"
print>>sys.stderr, '\nPARAMETERS:'
print>>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print>>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print>>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % args.min_hashsize
print>>sys.stderr, ' - paired = %s \t\t(-p)' % args.paired
print>>sys.stderr, ''
print>>sys.stderr, 'Estimated memory usage is %.2g bytes (n_hashes x min_hashsize)' % (args.n_hashes * args.min_hashsize)
print>>sys.stderr, '-'*8
K=args.ksize
HT_SIZE=args.min_hashsize
N_HT=args.n_hashes
DESIRED_COVERAGE=args.cutoff
report_fp = args.report_file
filenames = args.input_filenames
# In paired mode we read two records at a time
batch_size = 1
if args.paired:
batch_size = 2
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keep'
outfp = open(output_name, 'w')
n = -1
for n, batch in enumerate(batchwise(screed.open(input_filename), batch_size)):
if n > 0 and n % 100000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
if report_fp:
print>>report_fp, total, total - discarded, \
1. - (discarded / float(total))
report_fp.flush()
total += batch_size
# If in paired mode, check that the reads are properly interleaved
if args.paired:
if not validpair(batch[0], batch[1]):
print >>sys.stderr, 'Error: Improperly interleaved pairs %s %s' % (batch[0].name, batch[1].name)
sys.exit(-1)
# Emit the batch of reads if any read passes the filter
# and all reads are longer than K
passed_filter = False
passed_length = True
for record in batch:
if len(record.sequence) < K:
passed_length = False
continue
seq = record.sequence.replace('N', 'A')
med, _, _ = ht.get_median_count(seq)
if med < DESIRED_COVERAGE:
ht.consume(seq)
passed_filter = True
# Emit records if any passed
if passed_length and passed_filter:
for record in batch:
if hasattr(record,'accuracy'):
outfp.write('@%s\n%s\n+\n%s\n' % (record.name,
record.sequence,
record.accuracy))
else:
outfp.write('>%s\n%s\n' % (record.name, record.sequence))
else:
discarded += batch_size
if -1 < n:
print 'DONE with', input_filename, '; kept', total - discarded, 'of',\
total, 'or', int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
else: print 'SKIPPED empty file', input_filename
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
print >>sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main():
info('load-graph.py', ['graph'])
args = get_parser().parse_args()
report_on_config(args, hashtype='hashbits')
base = args.output_filename
filenames = args.input_filenames
n_threads = int(args.n_threads)
for _ in args.input_filenames:
check_file_status(_)
check_space(args.input_filenames)
check_space_for_hashtable(float(args.n_tables * args.min_tablesize) / 8.)
print 'Saving k-mer presence table to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
if args.no_build_tagset:
print 'We WILL NOT build the tagset.'
else:
print 'We WILL build the tagset (for partitioning/traversal).'
print 'making k-mer presence table'
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
if args.no_build_tagset:
target_method = htable.consume_fasta_with_reads_parser
else:
target_method = htable.consume_fasta_and_tag_with_reads_parser
config = khmer.get_config()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
for _, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input', filename
for _ in xrange(n_threads):
cur_thrd = threading.Thread(target=target_method, args=(rparser, ))
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
if args.report_total_kmers:
print >> sys.stderr, 'Total number of k-mers: {0}'.format(
htable.n_occupied())
print 'saving k-mer presence table in', base + '.pt'
htable.save(base + '.pt')
if not args.no_build_tagset:
print 'saving tagset in', base + '.tagset'
htable.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % htable.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
if args.write_fp_rate:
print >> info_fp, \
'\nfalse positive rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the graph structure is too small for "
"this data set. Increase table size/# tables.")
print >> sys.stderr, "**"
sys.exit(1)
def main():
info('filter-abund-single.py', ['counting', 'SeqAn'])
args = get_parser().parse_args()
check_input_files(args.datafile, args.force)
check_space([args.datafile], args.force)
if args.savetable:
check_space_for_hashtable(args.n_tables * args.min_tablesize,
args.force)
report_on_config(args)
print >> sys.stderr, 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables)
# first, load reads into hash table
rparser = khmer.ReadParser(args.datafile)
threads = []
print >> sys.stderr, 'consuming input, round 1 --', args.datafile
for _ in xrange(args.threads):
cur_thread = \
threading.Thread(
target=htable.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thread)
cur_thread.start()
for _ in threads:
_.join()
if args.report_total_kmers:
print >> sys.stderr, 'Total number of unique k-mers: {0}'.format(
htable.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(htable, args.force)
print >> sys.stderr, 'fp rate estimated to be %1.3f' % fp_rate
# now, trim.
# the filtering function.
def process_fn(record):
name = record.name
seq = record.sequence
seqN = seq.replace('N', 'A')
_, trim_at = htable.trim_on_abundance(seqN, args.cutoff)
if trim_at >= args.ksize:
# be sure to not to change the 'N's in the trimmed sequence -
# so, return 'seq' and not 'seqN'.
return name, seq[:trim_at]
return None, None
# the filtering loop
print >> sys.stderr, 'filtering', args.datafile
outfile = os.path.basename(args.datafile) + '.abundfilt'
outfp = open(outfile, 'w')
tsp = ThreadedSequenceProcessor(process_fn)
tsp.start(verbose_loader(args.datafile), outfp)
print >> sys.stderr, 'output in', outfile
if args.savetable:
print >>sys.stderr, 'Saving k-mer counting table filename', \
args.savetable
print >> sys.stderr, '...saving to', args.savetable
htable.save(args.savetable)
print >> sys.stderr, 'wrote to: ', outfile
def main(): # pylint: disable=too-many-locals,too-many-statements
info("do-partition.py", ["graph"])
args = sanitize_help(get_parser()).parse_args()
report_on_config(args, graphtype="nodegraph")
for infile in args.input_filenames:
check_input_files(infile, args.force)
check_space(args.input_filenames, args.force)
print("Saving k-mer nodegraph to %s" % args.graphbase, file=sys.stderr)
print("Loading kmers from sequences in %s" % repr(args.input_filenames), file=sys.stderr)
print("--", file=sys.stderr)
print("SUBSET SIZE", args.subset_size, file=sys.stderr)
print("N THREADS", args.threads, file=sys.stderr)
print("--", file=sys.stderr)
# load-graph.py
print("making nodegraph", file=sys.stderr)
nodegraph = khmer_args.create_nodegraph(args)
for _, filename in enumerate(args.input_filenames):
print("consuming input", filename, file=sys.stderr)
nodegraph.consume_fasta_and_tag(filename)
# 0.18 is ACTUAL MAX. Do not change.
fp_rate = khmer.calc_expected_collisions(nodegraph, args.force, max_false_pos=0.15)
print("fp rate estimated to be %1.3f" % fp_rate, file=sys.stderr)
# partition-graph
# do we want to exhaustively traverse the graph?
stop_big_traversals = args.no_big_traverse
if stop_big_traversals:
print("** This script brakes for lumps: ", "stop_big_traversals is true.", file=sys.stderr)
else:
print("** Traverse all the things:", " stop_big_traversals is false.", file=sys.stderr)
#
# now, partition!
#
# divide the tags up into subsets
divvy = nodegraph.divide_tags_into_subsets(int(args.subset_size))
divvy = list(divvy)
n_subsets = len(divvy)
divvy.append(0)
# build a queue of tasks:
worker_q = queue.Queue()
# break up the subsets into a list of worker tasks
for _ in range(0, n_subsets):
start = divvy[_]
end = divvy[_ + 1]
worker_q.put((nodegraph, _, start, end))
print("enqueued %d subset tasks" % n_subsets, file=sys.stderr)
open("%s.info" % args.graphbase, "w").write("%d subsets total\n" % (n_subsets))
if n_subsets < args.threads:
args.threads = n_subsets
# start threads!
print("starting %d threads" % args.threads, file=sys.stderr)
print("---", file=sys.stderr)
threads = []
for _ in range(args.threads):
cur_thread = threading.Thread(target=worker, args=(worker_q, args.graphbase, stop_big_traversals))
threads.append(cur_thread)
cur_thread.start()
assert threading.active_count() == args.threads + 1
print("done starting threads", file=sys.stderr)
# wait for threads
for _ in threads:
_.join()
print("---", file=sys.stderr)
print("done making subsets! see %s.subset.*.pmap" % (args.graphbase,), file=sys.stderr)
# merge-partitions
pmap_files = glob.glob(args.graphbase + ".subset.*.pmap")
print("loading %d pmap files (first one: %s)" % (len(pmap_files), pmap_files[0]), file=sys.stderr)
nodegraph = khmer.Nodegraph(args.ksize, 1, 1)
for pmap_file in pmap_files:
print("merging", pmap_file, file=sys.stderr)
nodegraph.merge_subset_from_disk(pmap_file)
if args.remove_subsets:
print("removing pmap files", file=sys.stderr)
for pmap_file in pmap_files:
os.unlink(pmap_file)
# annotate-partitions
for infile in args.input_filenames:
print("outputting partitions for", infile, file=sys.stderr)
outfile = os.path.basename(infile) + ".part"
part_count = nodegraph.output_partitions(infile, outfile)
print("output %d partitions for %s" % (part_count, infile), file=sys.stderr)
print("partitions are in", outfile, file=sys.stderr)
def main(): # pylint: disable=too-many-branches,too-many-statements
parser = sanitize_help(get_parser())
args = parser.parse_args()
configure_logging(args.quiet)
report_on_config(args)
report_fp = args.report
force_single = args.force_single
# check for similar filenames
# if we're using a single output file only check for identical filenames
# otherwise, check for identical BASE names as well.
filenames = []
basenames = []
for pathfilename in args.input_filenames:
filenames.append(pathfilename)
if args.single_output_file:
continue # nothing more to worry about
basename = os.path.basename(pathfilename)
if basename in basenames:
log_error('ERROR: Duplicate filename--Cannot handle this!')
log_error('** Exiting!')
sys.exit(1)
basenames.append(basename)
# check that files exist and there is sufficient output disk space.
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savegraph is not None:
graphsize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.savegraph, graphsize, args.force)
# load or create counting table.
if args.loadgraph:
log_info('loading k-mer countgraph from {graph}',
graph=args.loadgraph)
countgraph = Countgraph.load(args.loadgraph)
else:
log_info('making countgraph')
countgraph = khmer_args.create_countgraph(args)
# create an object to handle diginorm of all files
norm = Normalizer(args.cutoff, countgraph)
with_diagnostics = WithDiagnostics(norm, report_fp, args.report_frequency)
# make a list of all filenames and if they're paired or not;
# if we don't know if they're paired, default to allowing but not
# forcing pairing.
files = []
for element in filenames:
files.append([element, args.paired])
if args.unpaired_reads:
files.append([args.unpaired_reads, False])
corrupt_files = []
outfp = None
output_name = None
if args.single_output_file:
outfp = get_file_writer(args.single_output_file, args.gzip, args.bzip)
else:
if '-' in filenames or '/dev/stdin' in filenames:
print("Accepting input from stdin; output filename must "
"be provided with '-o'.", file=sys.stderr)
sys.exit(1)
#
# main loop: iterate over all files given, do diginorm.
#
for filename, require_paired in files:
if not args.single_output_file:
output_name = os.path.basename(filename) + '.keep'
outfp = open(output_name, 'wb')
outfp = get_file_writer(outfp, args.gzip, args.bzip)
# failsafe context manager in case an input file breaks
with catch_io_errors(filename, outfp, args.single_output_file,
args.force, corrupt_files):
screed_iter = clean_input_reads(screed.open(filename))
reader = broken_paired_reader(screed_iter, min_length=args.ksize,
force_single=force_single,
require_paired=require_paired)
# actually do diginorm
for record in with_diagnostics(reader, filename):
if record is not None:
write_record(record, outfp)
log_info('output in {name}', name=describe_file_handle(outfp))
if not args.single_output_file:
outfp.close()
# finished - print out some diagnostics.
log_info('Total number of unique k-mers: {umers}',
umers=countgraph.n_unique_kmers())
if args.savegraph is not None:
log_info('...saving to {name}', name=args.savegraph)
countgraph.save(args.savegraph)
fp_rate = \
khmer.calc_expected_collisions(countgraph, False, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
log_info('fp rate estimated to be {fpr:1.3f}', fpr=fp_rate)
if args.force and len(corrupt_files) > 0:
log_error("** WARNING: Finished with errors!")
log_error("** I/O Errors occurred in the following files:")
log_error("\t" + " ".join(corrupt_files))
def main():
info('correct-reads.py', ['streaming'])
args = sanitize_help(get_parser()).parse_args()
###
if len(set(args.input_filenames)) != len(args.input_filenames):
print("Error: Cannot input the same filename multiple times.",
file=sys.stderr)
sys.exit(1)
###
report_on_config(args)
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
tablesize = calculate_graphsize(args, 'countgraph')
if args.savegraph:
check_space_for_graph(args.savegraph, tablesize, args.force)
K = args.ksize
CUTOFF = args.cutoff
NORMALIZE_LIMIT = args.normalize_to
if args.loadgraph:
print('loading k-mer countgraph from', args.loadgraph, file=sys.stderr)
ct = Countgraph.load(args.loadgraph)
else:
print('making k-mer countgraph', file=sys.stderr)
ct = create_countgraph(args, multiplier=8 / (9. + 0.3))
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print('created temporary directory %s; use -T to change location' %
tempdir,
file=sys.stderr)
aligner = khmer.ReadAligner(ct, args.cutoff, args.bits_theta)
# ### FIRST PASS ###
save_pass2_total = 0
n_bp = 0
n_reads = 0
written_bp = 0
written_reads = 0
corrected_reads = 0
pass2list = []
for filename in args.input_filenames:
pass2filename = os.path.basename(filename) + '.pass2'
pass2filename = os.path.join(tempdir, pass2filename)
if args.out is None:
corrfp = open(os.path.basename(filename) + '.corr', 'w')
else:
corrfp = args.out
pass2list.append((filename, pass2filename, corrfp))
screed_iter = screed.open(filename, parse_description=False)
pass2fp = open(pass2filename, 'w')
save_pass2 = 0
n = 0
paired_iter = broken_paired_reader(screed_iter,
min_length=K,
force_single=args.ignore_pairs)
for n, is_pair, read1, read2 in paired_iter:
if n % 10000 == 0:
print('...',
n,
filename,
save_pass2,
n_reads,
n_bp,
written_reads,
written_bp,
file=sys.stderr)
# we want to track paired reads here, to make sure that pairs
# are not split between first pass and second pass.
if is_pair:
n_reads += 2
n_bp += len(read1.sequence) + len(read2.sequence)
seq1 = read1.sequence.replace('N', 'A')
seq2 = read2.sequence.replace('N', 'A')
med1, _, _ = ct.get_median_count(seq1)
med2, _, _ = ct.get_median_count(seq2)
if med1 < NORMALIZE_LIMIT or med2 < NORMALIZE_LIMIT:
ct.consume(seq1)
ct.consume(seq2)
write_record_pair(read1, read2, pass2fp)
save_pass2 += 2
else:
is_aligned, new_seq1 = correct_sequence(aligner, seq1)
if is_aligned:
if new_seq1 != read1.sequence:
corrected_reads += 1
read1.sequence = new_seq1
if hasattr(read1, 'quality'):
fix_quality(read1)
is_aligned, new_seq2 = correct_sequence(aligner, seq2)
if is_aligned:
if new_seq2 != read2.sequence:
corrected_reads += 1
read2.sequence = new_seq2
if hasattr(read2, 'quality'):
fix_quality(read2)
write_record_pair(read1, read2, corrfp)
written_reads += 2
written_bp += len(read1)
written_bp += len(read2)
else:
n_reads += 1
n_bp += len(read1.sequence)
seq = read1.sequence.replace('N', 'A')
med, _, _ = ct.get_median_count(seq)
# has this portion of the graph saturated? if not,
# consume & save => pass2.
if med < NORMALIZE_LIMIT:
ct.consume(seq)
write_record(read1, pass2fp)
save_pass2 += 1
else: # trim!!
is_aligned, new_seq = correct_sequence(aligner, seq)
if is_aligned:
if new_seq != read1.sequence:
corrected_reads += 1
read1.sequence = new_seq
if hasattr(read1, 'quality'):
fix_quality(read1)
write_record(read1, corrfp)
written_reads += 1
written_bp += len(new_seq)
pass2fp.close()
print('%s: kept aside %d of %d from first pass, in %s' %
(filename, save_pass2, n, filename),
file=sys.stderr)
save_pass2_total += save_pass2
# ### SECOND PASS. ###
skipped_n = 0
skipped_bp = 0
for _, pass2filename, corrfp in pass2list:
print(('second pass: looking at sequences kept aside in %s') %
pass2filename,
file=sys.stderr)
# note that for this second pass, we don't care about paired
# reads - they will be output in the same order they're read in,
# so pairs will stay together if not orphaned. This is in contrast
# to the first loop.
for n, read in enumerate(
screed.open(pass2filename, parse_description=False)):
if n % 10000 == 0:
print('... x 2',
n,
pass2filename,
written_reads,
written_bp,
file=sys.stderr)
seq = read.sequence.replace('N', 'A')
med, _, _ = ct.get_median_count(seq)
# do we retain low-abundance components unchanged?
if med < NORMALIZE_LIMIT and args.variable_coverage:
write_record(read, corrfp)
written_reads += 1
written_bp += len(read.sequence)
skipped_n += 1
skipped_bp += len(read.sequence)
# otherwise, examine/correct.
else: # med >= NORMALIZE LIMIT or not args.variable_coverage
is_aligned, new_seq = correct_sequence(aligner, seq)
if is_aligned:
if new_seq != read.sequence:
corrected_reads += 1
read.sequence = new_seq
if hasattr(read, 'quality'):
fix_quality(read)
write_record(read, corrfp)
written_reads += 1
written_bp += len(new_seq)
print('removing %s' % pass2filename, file=sys.stderr)
os.unlink(pass2filename)
print('removing temp directory & contents (%s)' % tempdir, file=sys.stderr)
shutil.rmtree(tempdir)
n_passes = 1.0 + (float(save_pass2_total) / n_reads)
percent_reads_corrected = float(corrected_reads +
(n_reads - written_reads)) /\
n_reads * 100.0
print('read %d reads, %d bp' % (
n_reads,
n_bp,
), file=sys.stderr)
print('wrote %d reads, %d bp' % (
written_reads,
written_bp,
),
file=sys.stderr)
print('looked at %d reads twice (%.2f passes)' %
(save_pass2_total, n_passes),
file=sys.stderr)
print('removed %d reads and corrected %d reads (%.2f%%)' %
(n_reads - written_reads, corrected_reads, percent_reads_corrected),
file=sys.stderr)
print('removed %.2f%% of bases (%d total)' %
((1 - (written_bp / float(n_bp))) * 100.0, n_bp - written_bp),
file=sys.stderr)
if args.variable_coverage:
percent_reads_hicov = 100.0 * float(n_reads - skipped_n) / n_reads
print('%d reads were high coverage (%.2f%%);' %
(n_reads - skipped_n, percent_reads_hicov),
file=sys.stderr)
print(('skipped %d reads/%d bases because of low coverage') %
(skipped_n, skipped_bp),
file=sys.stderr)
fp_rate = \
khmer.calc_expected_collisions(ct, args.force, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
print('fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate),
file=sys.stderr)
print('output in *.corr', file=sys.stderr)
if args.savegraph:
print("Saving k-mer countgraph to", args.savegraph, file=sys.stderr)
ct.save(args.savegraph)
def main():
info('trim-low-abund.py', ['streaming'])
parser = get_parser()
args = parser.parse_args()
###
if len(set(args.input_filenames)) != len(args.input_filenames):
print >>sys.stderr, \
"Error: Cannot input the same filename multiple times."
sys.exit(1)
###
report_on_config(args)
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savetable:
check_space_for_hashtable(args.n_tables * args.min_tablesize,
args.force)
K = args.ksize
CUTOFF = args.cutoff
NORMALIZE_LIMIT = args.normalize_to
if args.loadtable:
print >> sys.stderr, 'loading k-mer counting table from', args.loadtable
ct = khmer.load_counting_hash(args.loadtable)
else:
print >> sys.stderr, 'making k-mer counting table'
ct = khmer.new_counting_hash(K, args.min_tablesize, args.n_tables)
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print >>sys.stderr, 'created temporary directory %s; ' \
'use -T to change location' % tempdir
# ### FIRST PASS ###
save_pass2_total = 0
n_bp = 0
n_reads = 0
written_bp = 0
written_reads = 0
trimmed_reads = 0
pass2list = []
for filename in args.input_filenames:
pass2filename = os.path.basename(filename) + '.pass2'
pass2filename = os.path.join(tempdir, pass2filename)
if args.out is None:
trimfp = open(os.path.basename(filename) + '.abundtrim', 'w')
else:
trimfp = args.out
pass2list.append((filename, pass2filename, trimfp))
screed_iter = screed.open(filename, parse_description=False)
pass2fp = open(pass2filename, 'w')
save_pass2 = 0
n = 0
paired_iter = broken_paired_reader(screed_iter,
min_length=K,
force_single=args.ignore_pairs)
for n, is_pair, read1, read2 in paired_iter:
if n % 10000 == 0:
print >>sys.stderr, '...', n, filename, save_pass2, \
n_reads, n_bp, written_reads, written_bp
# we want to track paired reads here, to make sure that pairs
# are not split between first pass and second pass.
if is_pair:
n_reads += 2
n_bp += len(read1.sequence) + len(read2.sequence)
seq1 = read1.sequence.replace('N', 'A')
seq2 = read2.sequence.replace('N', 'A')
med1, _, _ = ct.get_median_count(seq1)
med2, _, _ = ct.get_median_count(seq2)
if med1 < NORMALIZE_LIMIT or med2 < NORMALIZE_LIMIT:
ct.consume(seq1)
ct.consume(seq2)
write_record_pair(read1, read2, pass2fp)
save_pass2 += 2
else:
_, trim_at1 = ct.trim_on_abundance(seq1, CUTOFF)
_, trim_at2 = ct.trim_on_abundance(seq2, CUTOFF)
if trim_at1 >= K:
read1 = trim_record(read1, trim_at1)
if trim_at2 >= K:
read2 = trim_record(read2, trim_at2)
if trim_at1 != len(seq1):
trimmed_reads += 1
if trim_at2 != len(seq2):
trimmed_reads += 1
write_record_pair(read1, read2, trimfp)
written_reads += 2
written_bp += trim_at1 + trim_at2
else:
n_reads += 1
n_bp += len(read1.sequence)
seq = read1.sequence.replace('N', 'A')
med, _, _ = ct.get_median_count(seq)
# has this portion of the graph saturated? if not,
# consume & save => pass2.
if med < NORMALIZE_LIMIT:
ct.consume(seq)
write_record(read1, pass2fp)
save_pass2 += 1
else: # trim!!
_, trim_at = ct.trim_on_abundance(seq, CUTOFF)
if trim_at >= K:
new_read = trim_record(read1, trim_at)
write_record(new_read, trimfp)
written_reads += 1
written_bp += trim_at
if trim_at != len(read1.sequence):
trimmed_reads += 1
pass2fp.close()
print >>sys.stderr, '%s: kept aside %d of %d from first pass, in %s' \
% (filename, save_pass2, n, filename)
save_pass2_total += save_pass2
# ### SECOND PASS. ###
skipped_n = 0
skipped_bp = 0
for _, pass2filename, trimfp in pass2list:
print >> sys.stderr, ('second pass: looking at sequences kept aside '
'in %s') % pass2filename
# note that for this second pass, we don't care about paired
# reads - they will be output in the same order they're read in,
# so pairs will stay together if not orphaned. This is in contrast
# to the first loop.
for n, read in enumerate(
screed.open(pass2filename, parse_description=False)):
if n % 10000 == 0:
print >>sys.stderr, '... x 2', n, pass2filename, \
written_reads, written_bp
seq = read.sequence.replace('N', 'A')
med, _, _ = ct.get_median_count(seq)
# do we retain low-abundance components unchanged?
if med < NORMALIZE_LIMIT and args.variable_coverage:
write_record(read, trimfp)
written_reads += 1
written_bp += len(read.sequence)
skipped_n += 1
skipped_bp += len(read.sequence)
# otherwise, examine/trim/truncate.
else: # med >= NORMALIZE LIMIT or not args.variable_coverage
_, trim_at = ct.trim_on_abundance(seq, CUTOFF)
if trim_at >= K:
new_read = trim_record(read, trim_at)
write_record(new_read, trimfp)
written_reads += 1
written_bp += trim_at
if trim_at != len(read.sequence):
trimmed_reads += 1
print >> sys.stderr, 'removing %s' % pass2filename
os.unlink(pass2filename)
print >> sys.stderr, 'removing temp directory & contents (%s)' % tempdir
shutil.rmtree(tempdir)
n_passes = 1.0 + (float(save_pass2_total) / n_reads)
percent_reads_trimmed = float(trimmed_reads + (n_reads - written_reads)) /\
n_reads * 100.0
print >> sys.stderr, 'read %d reads, %d bp' % (
n_reads,
n_bp,
)
print >> sys.stderr, 'wrote %d reads, %d bp' % (
written_reads,
written_bp,
)
print >>sys.stderr, 'looked at %d reads twice (%.2f passes)' % \
(save_pass2_total, n_passes)
print >>sys.stderr, 'removed %d reads and trimmed %d reads (%.2f%%)' % \
(n_reads - written_reads, trimmed_reads, percent_reads_trimmed)
print >>sys.stderr, 'trimmed or removed %.2f%% of bases (%d total)' % \
((1 - (written_bp / float(n_bp))) * 100.0, n_bp - written_bp)
if args.variable_coverage:
percent_reads_hicov = 100.0 * float(n_reads - skipped_n) / n_reads
print >>sys.stderr, '%d reads were high coverage (%.2f%%);' % \
(n_reads - skipped_n, percent_reads_hicov)
print >> sys.stderr, ('skipped %d reads/%d bases because of low'
'coverage') % (skipped_n, skipped_bp)
fp_rate = \
khmer.calc_expected_collisions(ct, args.force, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
print >>sys.stderr, \
'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
print >> sys.stderr, 'output in *.abundtrim'
if args.savetable:
print >> sys.stderr, "Saving k-mer counting table to", args.savetable
ct.save(args.savetable)
def main():
parser = build_construct_args()
parser.add_argument('input_filename')
parser.add_argument('read_filename')
args = parser.parse_args()
if not args.quiet:
if args.min_hashsize == DEFAULT_MIN_HASHSIZE:
print >>sys.stderr, "** WARNING: hashsize is default! " \
"You absodefly want to increase this!\n** " \
"Please read the docs!"
print >>sys.stderr, '\nPARAMETERS:'
print >>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % \
args.min_hashsize
print >>sys.stderr, ''
print >>sys.stderr, 'Estimated memory usage is %.2g bytes ' \
'(n_hashes x min_hashsize / 8)' % (
args.n_hashes * args.min_hashsize / 8.)
print >>sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
inp = args.input_filename
readsfile = args.read_filename
tag = args.tag
outfile = os.path.basename(readsfile) + '.' + tag + '.sweep2'
outfp = open(outfile, 'w')
# create a hashbits data structure
ht = khmer.new_hashbits(K, HT_SIZE, N_HT)
# load contigs, connect into N partitions
print 'loading input reads from', inp
ht.consume_fasta(inp)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
print >>sys.stderr, "** Do not use these results!!"
sys.exit(-1)
print 'starting sweep.'
n = 0
m = 0
for record in screed.open(readsfile):
if len(record.sequence) < K:
continue
if n % 100000 == 0:
print '...', n, m
count = ht.get_median_count(record.sequence)[0]
if count:
m += 1
outfp.write('>%s\n%s\n' % (record.name, record.sequence))
n += 1
def main():
info('load-into-counting.py', ['counting', 'SeqAn'])
args = get_parser().parse_args()
report_on_config(args)
base = args.output_countingtable_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_input_files(name, args.force)
check_space(args.input_sequence_filename, args.force)
check_space_for_hashtable(args.n_tables * args.min_tablesize, args.force)
check_file_writable(base)
check_file_writable(base + ".info")
print >>sys.stderr, 'Saving k-mer counting table to %s' % base
print >>sys.stderr, 'Loading kmers from sequences in %s' % repr(filenames)
# clobber the '.info' file now, as we always open in append mode below
if os.path.exists(base + '.info'):
os.remove(base + '.info')
print >>sys.stderr, 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables)
htable.set_use_bigcount(args.bigcount)
filename = None
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
print >>sys.stderr, 'consuming input', filename
for _ in xrange(args.threads):
cur_thrd = \
threading.Thread(
target=htable.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
if index > 0 and index % 10 == 0:
check_space_for_hashtable(args.n_tables * args.min_tablesize,
args.force)
print >>sys.stderr, 'mid-save', base
htable.save(base)
with open(base + '.info', 'a') as info_fh:
print >> info_fh, 'through', filename
n_kmers = htable.n_unique_kmers()
if args.report_total_kmers:
print >> sys.stderr, 'Total number of unique k-mers:', n_kmers
with open(base + '.info', 'a') as info_fp:
print >>info_fp, 'Total number of unique k-mers:', n_kmers
print >>sys.stderr, 'saving', base
htable.save(base)
# Change max_false_pos=0.2 only if you really grok it. HINT: You don't
fp_rate = \
khmer.calc_expected_collisions(htable, args.force, max_false_pos=.2)
with open(base + '.info', 'a') as info_fp:
print >> info_fp, 'fp rate estimated to be %1.3f\n' % fp_rate
if args.summary_info:
mr_fmt = args.summary_info.lower()
mr_file = base + '.info.' + mr_fmt
print >> sys.stderr, "Writing summmary info to", mr_file
with open(mr_file, 'w') as mr_fh:
if mr_fmt == 'json':
mr_data = {
"ht_name": os.path.basename(base),
"fpr": fp_rate,
"num_kmers": n_kmers,
"files": filenames,
"mrinfo_version": "0.1.0",
}
json.dump(mr_data, mr_fh)
mr_fh.write('\n')
elif mr_fmt == 'tsv':
mr_fh.write("ht_name\tfpr\tnum_kmers\tfiles\n")
mr_fh.write("{b:s}\t{fpr:1.3f}\t{k:d}\t{fls:s}\n".format(
b=os.path.basename(base), fpr=fp_rate, k=n_kmers,
fls=";".join(filenames)))
print >> sys.stderr, 'fp rate estimated to be %1.3f' % fp_rate
print >>sys.stderr, 'DONE.'
print >>sys.stderr, 'wrote to:', base + '.info'
def main(): # pylint: disable=too-many-branches,too-many-statements
info('normalize-by-median.py', ['diginorm'])
args = get_parser().parse_args()
report_on_config(args)
report_fp = args.report
# check for similar filenames
filenames = []
for pathfilename in args.input_filenames:
filename = pathfilename.split('/')[-1]
if (filename in filenames):
print >>sys.stderr, "WARNING: At least two input files are named \
%s . (The script normalize-by-median.py can not handle this, only one .keep \
file for one of the input files will be generated.)" % filename
else:
filenames.append(filename)
# check for others
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savetable:
check_space_for_hashtable(
args.n_tables * args.min_tablesize, args.force)
# list to save error files along with throwing exceptions
corrupt_files = []
if args.loadtable:
print 'loading k-mer counting table from', args.loadtable
htable = khmer.load_counting_hash(args.loadtable)
else:
print >> sys.stderr, 'making k-mer counting table'
htable = khmer.new_counting_hash(args.ksize, args.min_tablesize,
args.n_tables)
input_filename = None
for index, input_filename in enumerate(args.input_filenames):
total_acc, discarded_acc, corrupt_files = \
normalize_by_median_and_check(
input_filename, htable, args.single_output_file,
args.fail_save, args.paired, args.cutoff, args.force,
corrupt_files, report_fp)
if (args.dump_frequency > 0 and
index > 0 and index % args.dump_frequency == 0):
print 'Backup: Saving k-mer counting file through', input_filename
if args.savetable:
hashname = args.savetable
print '...saving to', hashname
else:
hashname = 'backup.ct'
print 'Nothing given for savetable, saving to', hashname
htable.save(hashname)
if args.paired and args.unpaired_reads:
args.paired = False
output_name = args.unpaired_reads
if not args.single_output_file:
output_name = os.path.basename(args.unpaired_reads) + '.keep'
outfp = open(output_name, 'w')
total_acc, discarded_acc, corrupt_files = \
normalize_by_median_and_check(
args.unpaired_reads, htable, args.single_output_file,
args.fail_save, args.paired, args.cutoff, args.force,
corrupt_files, report_fp)
if args.report_total_kmers:
print >> sys.stderr, 'Total number of unique k-mers: {0}'.format(
htable.n_unique_kmers())
if args.savetable:
print 'Saving k-mer counting table through', input_filename
print '...saving to', args.savetable
htable.save(args.savetable)
fp_rate = \
khmer.calc_expected_collisions(htable, args.force, max_false_pos=.8)
# for max_false_pos see Zhang et al., http://arxiv.org/abs/1309.2975
print >> sys.stderr, \
'fp rate estimated to be {fpr:1.3f}'.format(fpr=fp_rate)
if args.force and len(corrupt_files) > 0:
print >> sys.stderr, "** WARNING: Finished with errors!"
print >> sys.stderr, "** IOErrors occurred in the following files:"
print >> sys.stderr, "\t", " ".join(corrupt_files)
def main():
args = sanitize_help(get_parser()).parse_args()
if not args.quiet:
info('load-into-counting.py', ['counting', 'SeqAn'])
configure_logging(args.quiet)
report_on_config(args)
base = args.output_countgraph_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_input_files(name, args.force)
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.output_countgraph_filename, tablesize,
args.force)
info_filename = base + ".info"
check_file_writable(base)
check_file_writable(info_filename)
log_info('Saving k-mer countgraph to {base}', base=base)
log_info('Loading kmers from sequences in {filenames}',
filenames=repr(filenames))
# clobber the '.info' file now, as we always open in append mode below
with open(info_filename, 'w') as info_fp:
print('khmer version:', khmer.__version__, file=info_fp)
log_info('making countgraph')
countgraph = khmer_args.create_countgraph(args)
countgraph.set_use_bigcount(args.bigcount)
filename = None
total_num_reads = 0
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
log_info('consuming input {input}', input=filename)
for _ in range(args.threads):
cur_thrd = \
threading.Thread(
target=countgraph.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
if index > 0 and index % 10 == 0:
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(base, tablesize, args.force)
log_info('mid-save {base}', base=base)
countgraph.save(base)
with open(info_filename, 'a') as info_fh:
print('through', filename, file=info_fh)
total_num_reads += rparser.num_reads
n_kmers = countgraph.n_unique_kmers()
log_info('Total number of unique k-mers: {nk}', nk=n_kmers)
with open(info_filename, 'a') as info_fp:
print('Total number of unique k-mers:', n_kmers, file=info_fp)
log_info('saving {base}', base=base)
countgraph.save(base)
# Change max_false_pos=0.2 only if you really grok it. HINT: You don't
fp_rate = \
khmer.calc_expected_collisions(
countgraph, args.force, max_false_pos=.2)
with open(info_filename, 'a') as info_fp:
print('fp rate estimated to be %1.3f\n' % fp_rate, file=info_fp)
if args.summary_info:
mr_fmt = args.summary_info.lower()
mr_file = base + '.info.' + mr_fmt
log_info("Writing summmary info to {mr_file}", mr_file=mr_file)
with open(mr_file, 'w') as mr_fh:
if mr_fmt == 'json':
mr_data = {
"ht_name": os.path.basename(base),
"fpr": fp_rate,
"num_kmers": n_kmers,
"files": filenames,
"mrinfo_version": "0.2.0",
"num_reads": total_num_reads,
}
json.dump(mr_data, mr_fh)
mr_fh.write('\n')
elif mr_fmt == 'tsv':
mr_fh.write("ht_name\tfpr\tnum_kmers\tnum_reads\tfiles\n")
vals = [
os.path.basename(base),
"{:1.3f}".format(fp_rate),
str(n_kmers),
str(total_num_reads),
";".join(filenames),
]
mr_fh.write("\t".join(vals) + "\n")
log_info('fp rate estimated to be {fpr:1.3f}', fpr=fp_rate)
log_info('DONE.')
log_info('wrote to: {filename}', filename=info_filename)
def main(): # pylint: disable=too-many-locals,too-many-statements
info('do-partition.py', ['graph'])
args = get_parser().parse_args()
report_on_config(args, hashtype='hashbits')
for infile in args.input_filenames:
check_file_status(infile)
check_space(args.input_filenames)
print 'Saving k-mer presence table to %s' % args.graphbase
print 'Loading kmers from sequences in %s' % repr(args.input_filenames)
print '--'
print 'SUBSET SIZE', args.subset_size
print 'N THREADS', args.n_threads
print '--'
# load-graph
print 'making k-mer presence table'
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
for _, filename in enumerate(args.input_filenames):
print 'consuming input', filename
htable.consume_fasta_and_tag(filename)
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the graph structure is too small for"
" this data set. Increase k-mer presence table "
"size/num of tables.")
print >> sys.stderr, "**"
sys.exit(1)
# partition-graph
# do we want to exhaustively traverse the graph?
stop_big_traversals = args.no_big_traverse
if stop_big_traversals:
print '** This script brakes for lumps: stop_big_traversals is true.'
else:
print '** Traverse all the things: stop_big_traversals is false.'
#
# now, partition!
#
# divide the tags up into subsets
divvy = htable.divide_tags_into_subsets(int(args.subset_size))
n_subsets = len(divvy)
divvy.append(0)
# build a queue of tasks:
worker_q = Queue.Queue()
# break up the subsets into a list of worker tasks
for _ in range(0, n_subsets):
start = divvy[_]
end = divvy[_ + 1]
worker_q.put((htable, _, start, end))
print 'enqueued %d subset tasks' % n_subsets
open('%s.info' % args.graphbase, 'w').write('%d subsets total\n'
% (n_subsets))
if n_subsets < args.n_threads:
args.n_threads = n_subsets
# start threads!
print 'starting %d threads' % args.n_threads
print '---'
threads = []
for _ in range(args.n_threads):
cur_thread = threading.Thread(target=worker,
args=(worker_q, args.graphbase,
stop_big_traversals))
threads.append(cur_thread)
cur_thread.start()
print 'done starting threads'
# wait for threads
for _ in threads:
_.join()
print '---'
print 'done making subsets! see %s.subset.*.pmap' % (args.graphbase,)
# merge-partitions
pmap_files = glob.glob(args.graphbase + '.subset.*.pmap')
print 'loading %d pmap files (first one: %s)' % (len(pmap_files),
pmap_files[0])
htable = khmer.new_hashbits(args.ksize, 1, 1)
for pmap_file in pmap_files:
print 'merging', pmap_file
htable.merge_subset_from_disk(pmap_file)
if args.remove_subsets:
print 'removing pmap files'
for pmap_file in pmap_files:
os.unlink(pmap_file)
# annotate-partitions
for infile in args.input_filenames:
print 'outputting partitions for', infile
outfile = os.path.basename(infile) + '.part'
part_count = htable.output_partitions(infile, outfile)
print 'output %d partitions for %s' % (part_count, infile)
print 'partitions are in', outfile
def main():
parser = build_counting_args()
parser.add_argument("-t", "--trusted-cutoff", dest="trusted_cutoff", type=int, default=3)
parser.add_argument("--bits-theta", help="Tuning parameter controlling trade off of speed vs alignment sensitivity", default=1.0, type=float, dest="bits_theta")
parser.add_argument('-C', '--cutoff', type=int, dest='cutoff',
default=DEFAULT_MINIMUM_COVERAGE)
parser.add_argument('-s', '--savehash', dest='savehash', default='')
parser.add_argument('-l', '--loadhash', dest='loadhash',
default='')
parser.add_argument('--details-out', dest="details_out")
parser.add_argument('input_filenames', nargs='+')
args = parser.parse_args()
if not args.quiet:
print >>sys.stderr, '\nPARAMETERS:'
print >>sys.stderr, ' - kmer size = %d \t\t(-k)' % args.ksize
print >>sys.stderr, ' - n hashes = %d \t\t(-N)' % args.n_hashes
print >>sys.stderr, ' - min hashsize = %-5.2g \t(-x)' % \
args.min_hashsize
print >>sys.stderr, ''
print >>sys.stderr, 'Estimated memory usage is %.2g bytes ' \
'(n_hashes x min_hashsize)' % (
args.n_hashes * args.min_hashsize)
print >>sys.stderr, '-' * 8
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
DESIRED_COVERAGE = args.cutoff
filenames = args.input_filenames
if args.loadhash:
print 'loading hashtable from', args.loadhash
ht = khmer.load_counting_hash(args.loadhash)
else:
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT)
aligner = khmer.new_readaligner(ht, args.trusted_cutoff, args.bits_theta)
if args.details_out != None:
details_out = open(args.details_out, "w")
else:
details_out = None
total = 0
discarded = 0
for input_filename in filenames:
output_name = os.path.basename(input_filename) + '.keepalign'
outfp = open(output_name, 'w')
for n, record in enumerate(screed.open(input_filename)):
if n > 0 and n % 10000 == 0:
print '... kept', total - discarded, 'of', total, ', or', \
int(100. - discarded / float(total) * 100.), '%'
print '... in file', input_filename
total += 1
if len(record.sequence) < K:
continue
seq = record.sequence.upper().replace('N', 'A')
##
score, graph_alignment, read_alignment, truncated = aligner.align(record.sequence)
keep = False
if truncated:
keep = True
else:
if False:
graph_seq = graph_alignment.replace("-", "")
else:
graph_seq = ""
for i in range(len(graph_alignment)):
if graph_alignment[i] == "-":
graph_seq += read_alignment[i]
else:
graph_seq += graph_alignment[i]
mincount = ht.get_min_count(graph_seq)
keep = True
seq = graph_seq
#if mincount < DESIRED_COVERAGE:
# keep = True
# seq = graph_seq
#else:
# assert not keep
if details_out != None:
details_out.write("+{7}\t{0:0.2f}\t{3}\t{4}\nread: {6}\ngraph_aln: {1}\nread_aln: {2}\nstored_seq:{5}\n".format(score, graph_alignment, read_alignment, truncated, keep, seq, record.sequence, record.name))
if keep:
ht.consume(seq)
outfp.write('>%s\n%s\n' % (record.name, seq))
else:
discarded += 1
print 'DONE with', input_filename, '; kept', total - discarded, 'of',\
total, 'or', int(100. - discarded / float(total) * 100.), '%'
print 'output in', output_name
if args.savehash:
print 'Saving hashfile through', input_filename
print '...saving to', args.savehash
ht.save(args.savehash)
# Change 0.2 only if you really grok it. HINT: You don't.
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.20:
print >>sys.stderr, "**"
print >>sys.stderr, "** ERROR: the counting hash is too small for"
print >>sys.stderr, "** this data set. Increase hashsize/num ht."
print >>sys.stderr, "**"
print >>sys.stderr, "** Do not use these results!!"
sys.exit(-1)
def main():
parser = build_construct_args(
"Filter k-mers at the given abundance (inmem version).")
add_threading_args(parser)
parser.add_argument('--cutoff', '-C', dest='cutoff',
default=DEFAULT_CUTOFF, type=int,
help="Trim at k-mers below this abundance.")
parser.add_argument('--savehash', dest='savehash', default='')
parser.add_argument('datafile')
args = parser.parse_args()
report_on_config(args)
K = args.ksize
HT_SIZE = args.min_hashsize
N_HT = args.n_hashes
n_threads = int(args.n_threads)
config = khmer.get_config()
bufsz = config.get_reads_input_buffer_size()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
print 'making hashtable'
ht = khmer.new_counting_hash(K, HT_SIZE, N_HT, n_threads)
filename = args.datafile
### first, load reads into hash table
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input, round 1 --', filename
for tnum in xrange(n_threads):
t = \
threading.Thread(
target=ht.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(t)
t.start()
for t in threads:
t.join()
fp_rate = khmer.calc_expected_collisions(ht)
print 'fp rate estimated to be %1.3f' % fp_rate
### now, trim.
### the filtering function.
def process_fn(record):
name = record['name']
seq = record['sequence']
if 'N' in seq:
return None, None
trim_seq, trim_at = ht.trim_on_abundance(seq, args.cutoff)
if trim_at >= K:
return name, trim_seq
return None, None
### the filtering loop
print 'filtering', filename
outfile = os.path.basename(filename) + '.abundfilt'
outfp = open(outfile, 'w')
tsp = ThreadedSequenceProcessor(process_fn)
tsp.start(verbose_loader(filename), outfp)
print 'output in', outfile
if args.savehash:
print 'Saving hashfile', args.savehash
print '...saving to', args.savehash
ht.save(args.savehash)
def main():
info('load-graph.py', ['graph'])
args = get_parser().parse_args()
report_on_config(args, hashtype='hashbits')
base = args.output_filename
filenames = args.input_filenames
n_threads = int(args.n_threads)
for _ in args.input_filenames:
check_file_status(_)
check_space(args.input_filenames)
check_space_for_hashtable(float(args.n_tables * args.min_tablesize) / 8.)
print 'Saving k-mer presence table to %s' % base
print 'Loading kmers from sequences in %s' % repr(filenames)
if args.no_build_tagset:
print 'We WILL NOT build the tagset.'
else:
print 'We WILL build the tagset (for partitioning/traversal).'
print 'making k-mer presence table'
htable = khmer.new_hashbits(args.ksize, args.min_tablesize, args.n_tables)
if args.no_build_tagset:
target_method = htable.consume_fasta_with_reads_parser
else:
target_method = htable.consume_fasta_and_tag_with_reads_parser
config = khmer.get_config()
config.set_reads_input_buffer_size(n_threads * 64 * 1024)
for _, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename, n_threads)
threads = []
print 'consuming input', filename
for _ in xrange(n_threads):
cur_thrd = threading.Thread(target=target_method, args=(rparser, ))
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
print 'saving k-mer presence table in', base + '.pt'
htable.save(base + '.pt')
if not args.no_build_tagset:
print 'saving tagset in', base + '.tagset'
htable.save_tagset(base + '.tagset')
info_fp = open(base + '.info', 'w')
info_fp.write('%d unique k-mers' % htable.n_unique_kmers())
fp_rate = khmer.calc_expected_collisions(htable)
print 'fp rate estimated to be %1.3f' % fp_rate
if fp_rate > 0.15: # 0.18 is ACTUAL MAX. Do not change.
print >> sys.stderr, "**"
print >> sys.stderr, ("** ERROR: the graph structure is too small for "
"this data set. Increase table size/# tables.")
print >> sys.stderr, "**"
sys.exit(1)
def main():
args = sanitize_help(get_parser()).parse_args()
configure_logging(args.quiet)
report_on_config(args)
base = args.output_countgraph_filename
filenames = args.input_sequence_filename
for name in args.input_sequence_filename:
check_input_files(name, args.force)
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.output_countgraph_filename, tablesize,
args.force)
info_filename = base + ".info"
check_file_writable(base)
check_file_writable(info_filename)
log_info('Saving k-mer countgraph to {base}', base=base)
log_info('Loading kmers from sequences in {filenames}',
filenames=repr(filenames))
# clobber the '.info' file now, as we always open in append mode below
with open(info_filename, 'w') as info_fp:
print('khmer version:', khmer.__version__, file=info_fp)
log_info('making countgraph')
countgraph = khmer_args.create_countgraph(args)
filename = None
total_num_reads = 0
for index, filename in enumerate(filenames):
rparser = khmer.ReadParser(filename)
threads = []
log_info('consuming input {input}', input=filename)
for _ in range(args.threads):
cur_thrd = \
threading.Thread(
target=countgraph.consume_seqfile_with_reads_parser,
args=(rparser, )
)
threads.append(cur_thrd)
cur_thrd.start()
for thread in threads:
thread.join()
if index > 0 and index % 10 == 0:
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(base, tablesize, args.force)
log_info('mid-save {base}', base=base)
countgraph.save(base)
with open(info_filename, 'a') as info_fh:
print('through', filename, file=info_fh)
total_num_reads += rparser.num_reads
n_kmers = countgraph.n_unique_kmers()
log_info('Total number of unique k-mers: {nk}', nk=n_kmers)
with open(info_filename, 'a') as info_fp:
print('Total number of unique k-mers:', n_kmers, file=info_fp)
log_info('saving {base}', base=base)
countgraph.save(base)
# Change max_false_pos=0.2 only if you really grok it. HINT: You don't
fp_rate = \
khmer.calc_expected_collisions(
countgraph, args.force, max_false_pos=.2)
with open(info_filename, 'a') as info_fp:
print('fp rate estimated to be %1.3f\n' % fp_rate, file=info_fp)
if args.summary_info:
mr_fmt = args.summary_info.lower()
mr_file = base + '.info.' + mr_fmt
log_info("Writing summmary info to {mr_file}", mr_file=mr_file)
with open(mr_file, 'w') as mr_fh:
if mr_fmt == 'json':
mr_data = {
"ht_name": os.path.basename(base),
"fpr": fp_rate,
"num_kmers": n_kmers,
"files": filenames,
"mrinfo_version": "0.2.0",
"num_reads": total_num_reads,
}
json.dump(mr_data, mr_fh)
mr_fh.write('\n')
elif mr_fmt == 'tsv':
mr_fh.write("ht_name\tfpr\tnum_kmers\tnum_reads\tfiles\n")
vals = [
os.path.basename(base),
"{:1.3f}".format(fp_rate),
str(n_kmers),
str(total_num_reads),
";".join(filenames),
]
mr_fh.write("\t".join(vals) + "\n")
log_info('fp rate estimated to be {fpr:1.3f}', fpr=fp_rate)
log_info('DONE.')
log_info('wrote to: {filename}', filename=info_filename)
def main(args):
# Input and output files
outstream = kevlar.open(args.out, 'w')
writer = kevlar.vcf.VCFWriter(
outstream,
source='kevlar::call',
refr=args.refr,
)
writer.write_header()
# Contigs = query sequences
contigstream = kevlar.parse_partitioned_reads(
kevlar.parse_augmented_fastx(kevlar.open(args.queryseq, 'r')))
contigs_by_partition = load_contigs(contigstream)
gdnastream = kevlar.parse_partitioned_reads(
kevlar.reference.load_refr_cutouts(kevlar.open(args.targetseq, 'r')))
mask = None
if args.gen_mask:
message = 'generating mask of variant-spanning k-mers'
kevlar.plog('[kevlar::call]', message)
ntables = 4
buckets = args.mask_mem * _buckets_per_byte['nodegraph'] / ntables
mask = khmer.Nodetable(args.ksize, buckets, ntables)
progress_indicator = kevlar.ProgressIndicator(
'[kevlar::call] processed contigs/gDNAs for {counter} partitions',
interval=10,
breaks=[100, 1000, 10000],
)
for partid, gdnas in gdnastream:
progress_indicator.update()
if partid not in contigs_by_partition:
continue
contigs = contigs_by_partition[partid]
caller = call(
gdnas,
contigs,
partid,
match=args.match,
mismatch=args.mismatch,
gapopen=args.open,
gapextend=args.extend,
ksize=args.ksize,
refrfile=args.refr,
debug=args.debug,
mindist=5,
homopolyfilt=not args.no_homopoly_filter,
maxtargetlen=args.max_target_length,
)
for varcall in caller:
if args.gen_mask:
window = varcall.attribute('ALTWINDOW')
if window is not None and len(window) >= args.ksize:
mask.consume(window)
writer.write(varcall)
if args.gen_mask:
fpr = khmer.calc_expected_collisions(mask, max_false_pos=1.0)
if fpr > args.mask_max_fpr:
message = 'WARNING: mask FPR is {:.4f}'.format(fpr)
message += '; exceeds user-specified limit'
message += ' of {:.4f}'.format(args.mask_max_fpr)
kevlar.plog('[kevlar::call]', message)
mask.save(args.gen_mask)
