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):
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):
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():
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():
sys.argv = parse_humanfriendly_mem(sys.argv)
info('filter-abund-single.py', ['counting', 'SeqAn'])
args = sanitize_help(get_parser()).parse_args()
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)
print('making countgraph', file=sys.stderr)
graph = khmer_args.create_countgraph(args)
# first, load reads into graph
rparser = khmer.ReadParser(args.datafile)
threads = []
print('consuming input, round 1 --', args.datafile, file=sys.stderr)
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()
print('Total number of unique k-mers: {0}'.format(
graph.n_unique_kmers()), file=sys.stderr)
fp_rate = khmer.calc_expected_collisions(graph, args.force)
print('fp rate estimated to be %1.3f' % fp_rate, file=sys.stderr)
# now, trim.
# the filtering function.
def process_fn(record):
name = record.name
seq = record.sequence
seqN = seq.replace('N', 'A')
_, trim_at = graph.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('filtering', args.datafile, file=sys.stderr)
outfile = os.path.basename(args.datafile) + '.abundfilt'
outfile = open(outfile, 'wb')
outfp = get_file_writer(outfile, args.gzip, args.bzip)
tsp = ThreadedSequenceProcessor(process_fn)
tsp.start(verbose_loader(args.datafile), outfp)
print('output in', outfile.name, file=sys.stderr)
if args.savegraph:
print('Saving k-mer countgraph filename',
args.savegraph, file=sys.stderr)
graph.save(args.savegraph)
def main():
info('trim-low-abund.py', ['streaming'])
parser = sanitize_help(get_parser())
args = 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)
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:
print(
"Accepting input from stdin; output filename must "
"be provided with -o.",
file=sys.stderr)
sys.exit(1)
if args.loadgraph:
print('loading countgraph from', args.loadgraph, file=sys.stderr)
ct = khmer.load_countgraph(args.loadgraph)
else:
print('making countgraph', file=sys.stderr)
ct = khmer_args.create_countgraph(args)
K = ct.ksize()
CUTOFF = args.cutoff
NORMALIZE_LIMIT = args.normalize_to
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print('created temporary directory %s; '
'use -T to change location' % tempdir,
file=sys.stderr)
# ### 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.output is None:
trimfp = get_file_writer(
open(os.path.basename(filename) + '.abundtrim', 'wb'),
args.gzip, args.bzip)
else:
trimfp = get_file_writer(args.output, args.gzip, args.bzip)
pass2list.append((filename, pass2filename, trimfp))
screed_iter = screed.open(filename)
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:
_, 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('%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, trimfp 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)):
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, 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('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_trimmed = float(trimmed_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 trimmed %d reads (%.2f%%)' %
(n_reads - written_reads, trimmed_reads, percent_reads_trimmed),
file=sys.stderr)
print('trimmed or 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 *.abundtrim', file=sys.stderr)
if args.savegraph:
print("Saving k-mer countgraph to", args.savegraph, file=sys.stderr)
ct.save(args.savegraph)
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'])
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.savegraph:
check_space_for_graph(
args.n_tables * args.min_tablesize, args.force)
K = args.ksize
CUTOFF = args.cutoff
NORMALIZE_LIMIT = args.normalize_to
if args.loadgraph:
print >>sys.stderr, 'loading k-mer countgraph from', args.loadgraph
ct = khmer.load_countgraph(args.loadgraph)
else:
print >>sys.stderr, 'making k-mer countgraph'
ct = khmer.new_countgraph(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
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 >>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:
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 >>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, corrfp 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, 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 >>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_corrected = float(corrected_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 corrected %d reads (%.2f%%)' % \
(n_reads - written_reads, corrected_reads, percent_reads_corrected)
print >>sys.stderr, '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 *.corr'
if args.savegraph:
print >>sys.stderr, "Saving k-mer countgraph to", args.savegraph
ct.save(args.savegraph)
def main():
info('collect-reads.py', ['counting'])
args = 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, False)
check_space(args.input_sequence_filename, False)
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.output_countgraph_filename, tablesize,
False)
print('Saving k-mer countgraph to %s' % base)
print('Loading sequences from %s' % repr(filenames))
if args.output:
print('Outputting sequences to', args.output)
print('making countgraph', file=sys.stderr)
htable = khmer_args.create_countgraph(args)
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('Total number of k-mers: {0}'.format(
htable.n_occupied()), file=sys.stderr)
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, False,
max_false_pos=.2)
print('fp rate estimated to be %1.3f' % fp_rate)
print('fp rate estimated to be %1.3f' % fp_rate, file=info_fp)
print('DONE.')
def main():
args = sanitize_help(get_parser()).parse_args()
if not args.quiet:
info('filter-abund-single.py', ['counting', 'SeqAn'])
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():
info('collect-reads.py', ['counting'])
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, False)
check_space(args.input_sequence_filename, False)
tablesize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.output_countgraph_filename, tablesize, False)
print('Saving k-mer countgraph to %s' % base)
print('Loading sequences from %s' % repr(filenames))
if args.output:
print('Outputting sequences to', args.output)
print('making countgraph', file=sys.stderr)
htable = khmer_args.create_countgraph(args)
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('Total number of k-mers: {0}'.format(htable.n_occupied()),
file=sys.stderr)
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, False, max_false_pos=.2)
print('fp rate estimated to be %1.3f' % fp_rate)
print('fp rate estimated to be %1.3f' % fp_rate, file=info_fp)
print('DONE.')
def main(): # pylint: disable=too-many-locals,too-many-branches
args = sanitize_help(get_parser()).parse_args()
configure_logging(args.quiet)
report_on_config(args)
check_input_files(args.input_sequence_filename, args.force)
if args.savegraph:
graphsize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.savegraph, graphsize, args.force)
if (not args.squash_output
and os.path.exists(args.output_histogram_filename)):
log_error('ERROR: {output} exists; not squashing.',
output=args.output_histogram_filename)
sys.exit(1)
else:
hist_fp = open(args.output_histogram_filename, 'w')
hist_fp_csv = csv.writer(hist_fp)
# write headers:
hist_fp_csv.writerow(
['abundance', 'count', 'cumulative', 'cumulative_fraction'])
log_info('making countgraph')
countgraph = khmer_args.create_countgraph(args, multiplier=1.1)
countgraph.set_use_bigcount(args.bigcount)
log_info('building k-mer tracking graph')
tracking = khmer_args.create_nodegraph(args, multiplier=1.1)
log_info('kmer_size: {ksize}', ksize=countgraph.ksize())
log_info('k-mer countgraph sizes: {sizes}', sizes=countgraph.hashsizes())
log_info('outputting to {output}', output=args.output_histogram_filename)
# start loading
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
log_info('consuming input, round 1 -- {input}',
input=args.input_sequence_filename)
for _ in range(args.threads):
thread = \
threading.Thread(
target=countgraph.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
log_info('Total number of unique k-mers: {nk}',
nk=countgraph.n_unique_kmers())
abundance_lists = []
def __do_abundance_dist__(read_parser):
abundances = countgraph.abundance_distribution_with_reads_parser(
read_parser, tracking)
abundance_lists.append(abundances)
log_info('preparing hist from {seqfile}...',
seqfile=args.input_sequence_filename)
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
log_info('consuming input, round 2 -- {filename}',
filename=args.input_sequence_filename)
for _ in range(args.threads):
thread = \
threading.Thread(
target=__do_abundance_dist__,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
assert len(abundance_lists) == args.threads, len(abundance_lists)
abundance = {}
for abundance_list in abundance_lists:
for i, count in enumerate(abundance_list):
abundance[i] = abundance.get(i, 0) + count
total = sum(abundance.values())
if 0 == total:
log_error("ERROR: abundance distribution is uniformly zero; "
"nothing to report.")
log_error("\tPlease verify that the input files are valid.")
sys.exit(1)
sofar = 0
for _, i in sorted(abundance.items()):
if i == 0 and not args.output_zero:
continue
sofar += i
frac = sofar / float(total)
hist_fp_csv.writerow([_, i, sofar, round(frac, 3)])
if sofar == total:
break
if args.savegraph:
log_info('Saving k-mer countgraph to {savegraph}',
savegraph=args.savegraph)
countgraph.save(args.savegraph)
log_info('wrote to: {output}', output=args.output_histogram_filename)
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,
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 = sanitize_help(get_parser()).parse_args()
if not args.quiet:
info('filter-abund-single.py', ['counting', 'SeqAn'])
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)
# now, trim.
# the filtering function.
def process_fn(record):
name = record.name
seq = record.sequence
seqN = seq.replace('N', 'A')
_, trim_at = graph.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
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)
tsp = ThreadedSequenceProcessor(process_fn, verbose=not args.quiet)
tsp.start(verbose_loader(args.datafile), 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():
info('trim-low-abund.py', ['streaming'])
parser = sanitize_help(get_parser())
args = 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)
if args.trim_at_coverage != DEFAULT_TRIM_AT_COVERAGE and \
not args.variable_coverage:
print("Error: --trim-at-coverage/-Z given, but",
"--variable-coverage/-V not specified.",
file=sys.stderr)
sys.exit(1)
if args.diginorm_coverage != DEFAULT_DIGINORM_COVERAGE and \
not args.diginorm:
print("Error: --diginorm-coverage given, but",
"--diginorm not specified.",
file=sys.stderr)
sys.exit(1)
if args.diginorm and args.single_pass:
print("Error: --diginorm and --single-pass are incompatible!\n"
"You probably want to use normalize-by-median.py instead.",
file=sys.stderr)
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:
print("Accepting input from stdin; output filename must "
"be provided with -o.", file=sys.stderr)
sys.exit(1)
if args.loadgraph:
print('loading countgraph from', args.loadgraph, file=sys.stderr)
ct = khmer.load_countgraph(args.loadgraph)
else:
print('making countgraph', file=sys.stderr)
ct = khmer_args.create_countgraph(args)
K = ct.ksize()
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print('created temporary directory %s; '
'use -T to change location' % tempdir, file=sys.stderr)
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:
print('...', filename, trimmer.n_saved,
trimmer.n_reads, trimmer.n_bp,
written_reads, written_bp, file=sys.stderr)
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()
print('%s: kept aside %d of %d from first pass, in %s' %
(filename,
trimmer.n_saved - save_start, trimmer.n_reads - n_start,
filename),
file=sys.stderr)
# 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:
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. 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:
print('... x 2', trimmer.n_reads - n_start,
pass2filename, trimmer.n_saved,
trimmer.n_reads, trimmer.n_bp,
written_reads, written_bp, file=sys.stderr)
watermark += REPORT_EVERY_N_READS
write_record(read, trimfp)
written_reads += 1
written_bp += len(read)
print('removing %s' % pass2filename, file=sys.stderr)
os.unlink(pass2filename)
# if we created our own trimfps, close 'em.
if not args.output:
trimfp.close()
print('removing temp directory & contents (%s)' % tempdir, file=sys.stderr)
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
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 trimmed %d reads (%.2f%%)' %
(n_reads - written_reads, trimmed_reads, percent_reads_trimmed),
file=sys.stderr)
print('trimmed or 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 - n_skipped) / n_reads
print('%d reads were high coverage (%.2f%%);' % (n_reads - n_skipped,
percent_reads_hicov),
file=sys.stderr)
print('skipped %d reads/%d bases because of low coverage' %
(n_skipped, bp_skipped),
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 *.abundtrim', file=sys.stderr)
if args.savegraph:
print("Saving k-mer countgraph to",
args.savegraph, file=sys.stderr)
ct.save(args.savegraph)
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 = sanitize_help(get_parser())
args = parser.parse_args()
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)
###
graphtype = 'countgraph' if not args.small_count else 'smallcountgraph'
report_on_config(args, graphtype=graphtype)
check_valid_file_exists(args.input_filenames)
check_space(args.input_filenames, args.force)
if args.savegraph:
graphsize = calculate_graphsize(args, graphtype)
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, args.small_count)
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.
paired_iter = broken_paired_reader(ReadParser(filename), 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.
read_parser = ReadParser(pass2filename)
paired_iter = broken_paired_reader(read_parser,
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)
read_parser.close()
log_info('removing {pass2}', pass2=pass2filename)
os.unlink(pass2filename)
# if we created our own trimfps, close 'em.
if not args.output:
trimfp.close()
try:
log_info('removing temp directory & contents ({temp})', temp=tempdir)
shutil.rmtree(tempdir)
except OSError as oe:
log_info('WARNING: unable to remove {temp} (probably an NFS issue); '
'please remove manually', temp=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)
if args.output is None:
log_info('output in *.abundtrim')
elif args.output.name == 1:
log_info('output streamed to stdout')
elif args.output.name:
log_info('output in {}'.format(args.output.name))
if args.savegraph:
log_info("Saving k-mer countgraph to {graph}", graph=args.savegraph)
ct.save(args.savegraph)
if args.summary_info is not None:
# note that when streaming to stdout the name of args.output will
# be set to 1
if args.output is not None and args.output.name != 1:
base = args.output.name
# no explicit name or stdout stream -> use a default name
else:
base = 'trim-low-abund-{}'.format(
time.strftime("%Y-%m-%dT%H:%M:%S"))
info = {'fpr': fp_rate,
'reads': n_reads,
'basepairs': n_bp,
'reads_written': written_reads,
'basepairs_written': written_bp,
'reads_skipped': n_skipped,
'basepairs_skipped': bp_skipped,
'reads_removed': n_reads - written_reads,
'reads_trimmed': trimmed_reads,
'basepairs_removed_or_trimmed': n_bp - written_bp
}
store_provenance_info(info, fname=base, format=args.summary_info)
def main(): # pylint: disable=too-many-branches,too-many-statements
parser = sanitize_help(get_parser())
args = parser.parse_args()
if not args.quiet:
info('normalize-by-median.py', ['diginorm'])
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:
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 = 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:
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('trim-low-abund.py', ['streaming'])
parser = sanitize_help(get_parser())
args = 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)
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:
print("Accepting input from stdin; output filename must "
"be provided with -o.", file=sys.stderr)
sys.exit(1)
if args.loadgraph:
print('loading countgraph from', args.loadgraph, file=sys.stderr)
ct = khmer.load_countgraph(args.loadgraph)
else:
print('making countgraph', file=sys.stderr)
ct = khmer_args.create_countgraph(args)
K = ct.ksize()
CUTOFF = args.cutoff
NORMALIZE_LIMIT = args.normalize_to
tempdir = tempfile.mkdtemp('khmer', 'tmp', args.tempdir)
print('created temporary directory %s; '
'use -T to change location' % tempdir, file=sys.stderr)
# ### 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.output is None:
trimfp = get_file_writer(open(os.path.basename(filename) +
'.abundtrim', 'wb'),
args.gzip, args.bzip)
else:
trimfp = get_file_writer(args.output, args.gzip, args.bzip)
pass2list.append((filename, pass2filename, trimfp))
screed_iter = screed.open(filename)
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:
_, 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('%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, trimfp 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)):
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, 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('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_trimmed = float(trimmed_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 trimmed %d reads (%.2f%%)' %
(n_reads - written_reads, trimmed_reads, percent_reads_trimmed),
file=sys.stderr)
print('trimmed or 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 *.abundtrim', file=sys.stderr)
if args.savegraph:
print("Saving k-mer countgraph to",
args.savegraph, file=sys.stderr)
ct.save(args.savegraph)
def main(): # pylint: disable=too-many-locals,too-many-branches
args = sanitize_help(get_parser()).parse_args()
graph_type = 'smallcountgraph' if args.small_count else 'countgraph'
configure_logging(args.quiet)
report_on_config(args, graph_type)
check_input_files(args.input_sequence_filename, args.force)
if args.savegraph is not None:
graphsize = calculate_graphsize(args, graph_type)
check_space_for_graph(args.savegraph, graphsize, args.force)
if (not args.squash_output and
os.path.exists(args.output_histogram_filename)):
log_error('ERROR: {output} exists; not squashing.',
output=args.output_histogram_filename)
sys.exit(1)
else:
hist_fp = open(args.output_histogram_filename, 'w')
hist_fp_csv = csv.writer(hist_fp)
# write headers:
hist_fp_csv.writerow(['abundance', 'count', 'cumulative',
'cumulative_fraction'])
log_info('making countgraph')
# In case the user specified a maximum memory usage, use 8/(9+eps) of that
# for the countgraph and 1/(9+eps) for the tracking nodegraph
# `eps` is used to account for the memory used by the python interpreter
countgraph = khmer_args.create_countgraph(args, multiplier=8 / (9. + 0.3))
log_info('building k-mer tracking graph')
tracking = khmer_args.create_matching_nodegraph(countgraph)
log_info('kmer_size: {ksize}', ksize=countgraph.ksize())
log_info('k-mer countgraph sizes: {sizes}', sizes=countgraph.hashsizes())
log_info('outputting to {output}', output=args.output_histogram_filename)
# start loading
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
log_info('consuming input, round 1 -- {input}',
input=args.input_sequence_filename)
for _ in range(args.threads):
thread = \
threading.Thread(
target=countgraph.consume_seqfile,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
log_info('Total number of unique k-mers: {nk}',
nk=countgraph.n_unique_kmers())
abundance_lists = []
def __do_abundance_dist__(read_parser):
abundances = countgraph.abundance_distribution(
read_parser, tracking)
abundance_lists.append(abundances)
log_info('preparing hist from {seqfile}...',
seqfile=args.input_sequence_filename)
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
log_info('consuming input, round 2 -- {filename}',
filename=args.input_sequence_filename)
for _ in range(args.threads):
thread = \
threading.Thread(
target=__do_abundance_dist__,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
assert len(abundance_lists) == args.threads, len(abundance_lists)
abundance = {}
for abundance_list in abundance_lists:
for i, count in enumerate(abundance_list):
abundance[i] = abundance.get(i, 0) + count
total = sum(abundance.values())
if 0 == total:
log_error("ERROR: abundance distribution is uniformly zero; "
"nothing to report.")
log_error("\tPlease verify that the input files are valid.")
sys.exit(1)
sofar = 0
for _, i in sorted(abundance.items()):
if i == 0 and not args.output_zero:
continue
sofar += i
frac = sofar / float(total)
hist_fp_csv.writerow([_, i, sofar, round(frac, 3)])
if sofar == total:
break
if args.savegraph is not None:
log_info('Saving k-mer countgraph to {savegraph}',
savegraph=args.savegraph)
countgraph.save(args.savegraph)
log_info('wrote to: {output}', output=args.output_histogram_filename)
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(): # pylint: disable=too-many-locals,too-many-branches
info('abundance-dist-single.py', ['counting', 'SeqAn'])
args = sanitize_help(get_parser()).parse_args()
report_on_config(args)
check_input_files(args.input_sequence_filename, args.force)
if args.savegraph:
graphsize = calculate_graphsize(args, 'countgraph')
check_space_for_graph(args.savegraph, graphsize, args.force)
if (not args.squash_output and
os.path.exists(args.output_histogram_filename)):
print('ERROR: %s exists; not squashing.' %
args.output_histogram_filename, file=sys.stderr)
sys.exit(1)
else:
hist_fp = open(args.output_histogram_filename, 'w')
hist_fp_csv = csv.writer(hist_fp)
# write headers:
hist_fp_csv.writerow(['abundance', 'count', 'cumulative',
'cumulative_fraction'])
print('making countgraph', file=sys.stderr)
countgraph = khmer_args.create_countgraph(args, multiplier=1.1)
countgraph.set_use_bigcount(args.bigcount)
print('building k-mer tracking graph', file=sys.stderr)
tracking = khmer_args.create_nodegraph(args, multiplier=1.1)
print('kmer_size:', countgraph.ksize(), file=sys.stderr)
print('k-mer countgraph sizes:',
countgraph.hashsizes(), file=sys.stderr)
print('outputting to', args.output_histogram_filename, file=sys.stderr)
# start loading
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
print('consuming input, round 1 --',
args.input_sequence_filename, file=sys.stderr)
for _ in range(args.threads):
thread = \
threading.Thread(
target=countgraph.consume_fasta_with_reads_parser,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
print('Total number of unique k-mers: {0}'.format(
countgraph.n_unique_kmers()), file=sys.stderr)
abundance_lists = []
def __do_abundance_dist__(read_parser):
abundances = countgraph.abundance_distribution_with_reads_parser(
read_parser, tracking)
abundance_lists.append(abundances)
print('preparing hist from %s...' %
args.input_sequence_filename, file=sys.stderr)
rparser = khmer.ReadParser(args.input_sequence_filename)
threads = []
print('consuming input, round 2 --',
args.input_sequence_filename, file=sys.stderr)
for _ in range(args.threads):
thread = \
threading.Thread(
target=__do_abundance_dist__,
args=(rparser, )
)
threads.append(thread)
thread.start()
for thread in threads:
thread.join()
assert len(abundance_lists) == args.threads, len(abundance_lists)
abundance = {}
for abundance_list in abundance_lists:
for i, count in enumerate(abundance_list):
abundance[i] = abundance.get(i, 0) + count
total = sum(abundance.values())
if 0 == total:
print("ERROR: abundance distribution is uniformly zero; "
"nothing to report.", file=sys.stderr)
print(
"\tPlease verify that the input files are valid.", file=sys.stderr)
sys.exit(1)
sofar = 0
for _, i in sorted(abundance.items()):
if i == 0 and not args.output_zero:
continue
sofar += i
frac = sofar / float(total)
hist_fp_csv.writerow([_, i, sofar, round(frac, 3)])
if sofar == total:
break
if args.savegraph:
print('Saving k-mer countgraph ', args.savegraph, file=sys.stderr)
print('...saving to', args.savegraph, file=sys.stderr)
countgraph.save(args.savegraph)
print('wrote to: ' + args.output_histogram_filename, file=sys.stderr)
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():
info("collect-reads.py", ["counting"])
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, False)
check_space(args.input_sequence_filename, False)
tablesize = calculate_graphsize(args, "countgraph")
check_space_for_graph(args.output_countgraph_filename, tablesize, False)
print("Saving k-mer countgraph to %s" % base)
print("Loading sequences from %s" % repr(filenames))
if args.output:
print("Outputting sequences to", args.output)
print("making countgraph", file=sys.stderr)
htable = khmer_args.create_countgraph(args)
htable.set_use_bigcount(args.bigcount)
total_coverage = 0.0
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("Total number of k-mers: {0}".format(htable.n_occupied()), file=sys.stderr)
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, False, max_false_pos=0.2)
print("fp rate estimated to be %1.3f" % fp_rate)
print("fp rate estimated to be %1.3f" % fp_rate, file=info_fp)
print("DONE.")