def run(self):
workspace = working_directory.Working(self.output_dir)
workspace.setup_reference(self.reference)
workspace.update_param(snp_cost=self.snp_cost)
# assert os.path.exists(self.reference), 'Reference file does not exist'
# reference_filename = workspace._object_filename('reference.fa')
# if os.path.exists(reference_filename):
# os.unlink(reference_filename)
# os.symlink(os.path.relpath(self.reference, self.output_dir), reference_filename)
bam_filename = io.abspath(self.output_dir, "alignments.bam")
bam_prefix = io.abspath(self.output_dir, "alignments")
if sam.is_bam(self.input):
sort_input_filename = self.input
temp_filename = None
else:
temp_filename = io.abspath(self.output_dir, "temp.bam")
sort_input_filename = temp_filename
writer = io.Pipe_writer(temp_filename, ["samtools", "view", "-S", "-b", "-"])
f = open(self.input, "rb")
while True:
data = f.read(1 << 20)
if not data:
break
writer.write(data)
writer.close()
f.close()
grace.status("Sort")
# io.execute([
# 'samtools', 'sort', '-n', sort_input_filename, bam_prefix
# ])
sam.sort_bam(sort_input_filename, bam_prefix, by_name=True)
if temp_filename is not None:
os.unlink(temp_filename)
grace.status("")
def run(self):
workspace = working_directory.Working(self.output_dir)
workspace.setup_reference(self.reference)
workspace.update_param(snp_cost = self.snp_cost)
#assert os.path.exists(self.reference), 'Reference file does not exist'
#reference_filename = workspace._object_filename('reference.fa')
#if os.path.exists(reference_filename):
# os.unlink(reference_filename)
#os.symlink(os.path.relpath(self.reference, self.output_dir), reference_filename)
bam_filename = io.abspath(self.output_dir, 'alignments.bam')
bam_prefix = io.abspath(self.output_dir, 'alignments')
if sam.is_bam(self.input):
sort_input_filename = self.input
temp_filename = None
else:
temp_filename = io.abspath(self.output_dir, 'temp.bam')
sort_input_filename = temp_filename
writer = io.Pipe_writer(temp_filename, ['samtools', 'view', '-S', '-b', '-'])
f = open(self.input, 'rb')
while True:
data = f.read(1<<20)
if not data: break
writer.write(data)
writer.close()
f.close()
grace.status('Sort')
#io.execute([
# 'samtools', 'sort', '-n', sort_input_filename, bam_prefix
#])
sam.sort_bam(sort_input_filename, bam_prefix, by_name=True)
if temp_filename is not None:
os.unlink(temp_filename)
grace.status('')
def run(self):
assert self.reads or self.pairs or self.interleaved, 'No reads given'
io.check_name_uniqueness(self.reads, self.pairs, self.interleaved)
working = self.get_workspace()
working.setup_reference(self.references, bowtie=True)
working.update_param(snp_cost=2.0)
reference = working.get_reference()
log_file = open(self.log_filename(),'wb')
with workspace.tempspace(dir=working.working_dir) as temp:
n = [ 0 ]
def tempname():
n[0] += 1
return temp/('%d.fq'%n[0])
def convert(filename):
info = io.get_file_info(filename)
ok = selection.matches('type-fastq:[compression-none/compression-gzip/compression-bzip2]', info)
if ok:
return filename
result_name = tempname()
with open(result_name,'wb') as f:
for name, seq, qual in io.read_sequences(filename, qualities='required'):
io.write_fastq(f, name, seq, qual)
return result_name
ones = [ ]
twos = [ ]
singles = [ ]
for pair in self.pairs:
assert len(pair) == 2, 'Need two files in each "pair:" section.'
ones.append(convert(pair[0]))
twos.append(convert(pair[1]))
for item in self.interleaved:
left_name = tempname()
right_name = tempname()
ones.append(left_name)
twos.append(right_name)
with open(left_name,'wb') as left, \
open(right_name,'wb') as right:
reader = io.read_sequences(item, qualities='required')
while True:
try:
name, seq, qual = reader.next()
except StopIteration:
break
io.write_fastq(left, name,seq,qual)
try:
name, seq, qual = reader.next()
except StopIteration:
raise grace.Error('Interleaved file contains odd number of sequences')
io.write_fastq(right, name,seq,qual)
for item in self.reads:
singles.append(convert(item))
cores = min(self.cores, legion.coordinator().get_cores())
command = (
[ 'bowtie2',
'--threads', str(cores),
'--rg-id', '1',
'--rg', 'SM:'+working.name,
] +
self.bowtie_options +
[ '-x', reference.get_bowtie_index_prefix() ]
)
commands = [ ]
if ones:
commands.append(command + [ '-1', ','.join(ones), '-2', ','.join(twos) ])
if singles:
commands.append(command + [ '-U', ','.join(singles) ])
temp_bam_name = temp/'temp.bam'
with io.pipe_to(
['samtools', 'view', '-S', '-b', '-'],
stdout=open(temp_bam_name,'wb'),
stderr=log_file
) as f:
header_sent = False
for command in commands:
self.log.log('Running:\n' + ' '.join(command) + '\n')
with io.pipe_from(
command,
stderr=log_file,
cores=cores
) as f_out:
for line in f_out:
if not header_sent or not line.startswith('@'):
f.write(line)
header_sent = True
#io.execute([
# 'samtools', 'sort', '-n', temp_bam_name, working/'alignments'
# ])
sam.sort_bam(temp_bam_name, working/'alignments', by_name=True, cores=self.cores)
log_file.close()
def run(self):
assert self.reads or self.pairs or self.interleaved, 'No reads given'
io.check_name_uniqueness(self.reads, self.pairs, self.interleaved)
working = self.get_workspace()
working.setup_reference(self.references, bowtie=True)
working.update_param(snp_cost=2.0)
reference = working.get_reference()
log_file = open(self.log_filename(), 'wb')
with workspace.tempspace(dir=working.working_dir) as temp:
n = [0]
def tempname():
n[0] += 1
return temp / ('%d.fq' % n[0])
def convert(filename):
info = io.get_file_info(filename)
ok = selection.matches(
'type-fastq:[compression-none/compression-gzip/compression-bzip2]',
info)
if ok:
return filename
result_name = tempname()
with open(result_name, 'wb') as f:
for name, seq, qual in io.read_sequences(
filename, qualities='required'):
io.write_fastq(f, name, seq, qual)
return result_name
ones = []
twos = []
singles = []
for pair in self.pairs:
assert len(
pair) == 2, 'Need two files in each "pair:" section.'
ones.append(convert(pair[0]))
twos.append(convert(pair[1]))
for item in self.interleaved:
left_name = tempname()
right_name = tempname()
ones.append(left_name)
twos.append(right_name)
with open(left_name,'wb') as left, \
open(right_name,'wb') as right:
reader = io.read_sequences(item, qualities='required')
while True:
try:
name, seq, qual = reader.next()
except StopIteration:
break
io.write_fastq(left, name, seq, qual)
try:
name, seq, qual = reader.next()
except StopIteration:
raise grace.Error(
'Interleaved file contains odd number of sequences'
)
io.write_fastq(right, name, seq, qual)
for item in self.reads:
singles.append(convert(item))
cores = min(self.cores, legion.coordinator().get_cores())
command = ([
'bowtie2',
'--threads',
str(cores),
'--rg-id',
'1',
'--rg',
'SM:' + working.name,
] + self.bowtie_options +
['-x', reference.get_bowtie_index_prefix()])
commands = []
if ones:
commands.append(command +
['-1', ','.join(ones), '-2', ','.join(twos)])
if singles:
commands.append(command + ['-U', ','.join(singles)])
temp_bam_name = temp / 'temp.bam'
with io.pipe_to(['samtools', 'view', '-S', '-b', '-'],
stdout=open(temp_bam_name, 'wb'),
stderr=log_file) as f:
header_sent = False
for command in commands:
self.log.log('Running:\n' + ' '.join(command) + '\n')
with io.pipe_from(command, stderr=log_file,
cores=cores) as f_out:
for line in f_out:
if not header_sent or not line.startswith('@'):
f.write(line)
header_sent = True
#io.execute([
# 'samtools', 'sort', '-n', temp_bam_name, working/'alignments'
# ])
sam.sort_bam(temp_bam_name,
working / 'alignments',
by_name=True,
cores=self.cores)
log_file.close()
def run(self):
grace.require_shrimp_2()
grace.require_samtools()
assert self.references, 'No reference sequences given'
assert self.reads or self.pairs or self.interleaved, 'No reads given'
for pair in self.pairs:
assert len(pair) == 2, 'Two files required in each pair: section'
io.check_name_uniqueness(self.reads, self.pairs, self.interleaved)
read_sets = []
for item in self.reads:
read_sets.append(([item], False))
for item in self.pairs:
read_sets.append((item, True))
for item in self.interleaved:
read_sets.append(([item], True))
#Create working directory
workspace = self.get_workspace()
workspace.setup_reference(self.references)
workspace.update_param(snp_cost=25)
reference = workspace.get_reference()
reference_filename = reference.reference_fasta_filename()
cores = min(self.cores, legion.coordinator().get_cores())
default_options = {
'-E': None,
'-T': None,
'-N': str(cores),
'-n': '2',
'-w': '200%',
'-p': 'opp-in',
'-I': '0,500',
'-X': None,
}
if self.sam_unaligned:
default_options['--sam-unaligned'] = None
if self.half_paired:
default_options['--half-paired'] = None
else:
default_options['--no-half-paired'] = None
cutoff = '55%' #Default changed in SHRiMP 2.0.2
if '-h' in self.shrimp_options:
cutoff = self.shrimp_options[self.shrimp_options.index('-h') + 1]
#Run shrimp
bam_filename = io.abspath(self.output_dir, 'alignments.bam')
bam_prefix = io.abspath(self.output_dir, 'alignments')
bam_sorted_prefix = io.abspath(self.output_dir, 'alignments_sorted')
temp_filename = io.abspath(self.output_dir, 'temp.bam')
log_filename = io.abspath(self.output_dir, 'shrimp_log.txt')
log_file = open(log_filename, 'wb')
sam_eater = sam.Bam_writer(temp_filename)
sam_header_sent = [False]
n_seen = [0]
def eat(f):
for line in f:
if line.startswith('@'):
if sam_header_sent[0]: continue
else:
n_seen[0] += 1
if n_seen[0] % 100000 == 0:
grace.status('%s alignments produced' %
grace.pretty_number(n_seen[0]))
sam_eater.write_raw(line)
sam_header_sent[0] = True
def remove_pair_options(options):
for flag in ['-p', '-I']:
while flag in options:
pos = options.index(flag)
options = options[:pos] + options[pos + 2:]
for flag in ['--half-paired']:
while flag in options:
pos = options.index(flag)
options = options[:pos] + options[pos + 1:]
return options
for i, (filenames, is_paired) in enumerate(read_sets):
options = self.shrimp_options[:]
has_qualities = all(
len(io.read_sequences(filename, qualities=True).next()) ==
3 #A little ugly
for filename in filenames)
if has_qualities:
options.append('--fastq')
if len(filenames) == 1:
reads_parameters = [filenames[0]]
else:
reads_parameters = ['-1', filenames[0], '-2', filenames[1]]
if '--qv-offset' not in self.shrimp_options:
#guesses = [ ]
#for filename in filenames:
# guesses.append(io.guess_quality_offset(filename))
#assert len(set(guesses)) == 1, 'Conflicting quality offset guesses, please specify --qv-offset manually.'
#default_options['--qv-offset'] = str(guesses[0])
default_options['--qv-offset'] = str(
io.guess_quality_offset(*filenames))
default_options['--read-group'] = '%s,%s' % (
workspace.name.replace(',',
'_'), workspace.name.replace(',', '_'))
for flag in default_options:
if flag not in options:
options.append(flag)
if default_options[flag] is not None:
options.append(default_options[flag])
if not is_paired:
options = remove_pair_options(options)
grace.status('')
full_param = reference.shrimp_command(self.cs,
options + reads_parameters)
print >> sys.stderr, 'Running', ' '.join(full_param)
with io.pipe_from(full_param, stderr=log_file, cores=cores) as f:
eat(f)
log_file.close()
sam_eater.close()
grace.status('Sort')
#io.execute([
# 'samtools', 'sort', '-n', temp_filename, bam_prefix
#])
sam.sort_bam(temp_filename, bam_prefix, by_name=True, cores=self.cores)
os.unlink(temp_filename)
grace.status('')
def run(self):
grace.require_shrimp_2()
grace.require_samtools()
assert self.references, 'No reference sequences given'
assert self.reads or self.pairs or self.interleaved, 'No reads given'
for pair in self.pairs:
assert len(pair) == 2, 'Two files required in each pair: section'
io.check_name_uniqueness(self.reads, self.pairs, self.interleaved)
read_sets = [ ]
for item in self.reads:
read_sets.append( ([item], False) )
for item in self.pairs:
read_sets.append( (item, True) )
for item in self.interleaved:
read_sets.append( ([item], True) )
#Create working directory
workspace = self.get_workspace()
workspace.setup_reference(self.references)
workspace.update_param(snp_cost=25)
reference = workspace.get_reference()
reference_filename = reference.reference_fasta_filename()
cores = min(self.cores, legion.coordinator().get_cores())
default_options = {
'-E' : None,
'-T' : None,
'-N' : str(cores),
'-n':'2',
'-w':'200%',
'-p': 'opp-in',
'-I': '0,500',
'-X':None,
}
if self.sam_unaligned:
default_options['--sam-unaligned'] = None
if self.half_paired:
default_options['--half-paired'] = None
else:
default_options['--no-half-paired'] = None
cutoff = '55%' #Default changed in SHRiMP 2.0.2
if '-h' in self.shrimp_options:
cutoff = self.shrimp_options[ self.shrimp_options.index('-h')+1 ]
#Run shrimp
bam_filename = io.abspath(self.output_dir, 'alignments.bam')
bam_prefix = io.abspath(self.output_dir, 'alignments')
bam_sorted_prefix = io.abspath(self.output_dir, 'alignments_sorted')
temp_filename = io.abspath(self.output_dir, 'temp.bam')
log_filename = io.abspath(self.output_dir, 'shrimp_log.txt')
log_file = open(log_filename, 'wb')
sam_eater = sam.Bam_writer(temp_filename)
sam_header_sent = [False]
n_seen = [0]
def eat(f):
for line in f:
if line.startswith('@'):
if sam_header_sent[0]: continue
else:
n_seen[0] += 1
if n_seen[0] % 100000 == 0:
grace.status('%s alignments produced' % grace.pretty_number(n_seen[0]))
sam_eater.write_raw(line)
sam_header_sent[0] = True
def remove_pair_options(options):
for flag in ['-p','-I']:
while flag in options:
pos = options.index(flag)
options = options[:pos] + options[pos+2:]
for flag in ['--half-paired']:
while flag in options:
pos = options.index(flag)
options = options[:pos] + options[pos+1:]
return options
for i, (filenames, is_paired) in enumerate(read_sets):
options = self.shrimp_options[:]
has_qualities = all(
len( io.read_sequences(filename, qualities=True).next() ) == 3 #A little ugly
for filename in filenames
)
if has_qualities:
options.append( '--fastq' )
if len(filenames) == 1:
reads_parameters = [ filenames[0] ]
else:
reads_parameters = [ '-1', filenames[0], '-2', filenames[1] ]
if '--qv-offset' not in self.shrimp_options:
#guesses = [ ]
#for filename in filenames:
# guesses.append(io.guess_quality_offset(filename))
#assert len(set(guesses)) == 1, 'Conflicting quality offset guesses, please specify --qv-offset manually.'
#default_options['--qv-offset'] = str(guesses[0])
default_options['--qv-offset'] = str( io.guess_quality_offset(*filenames) )
default_options['--read-group'] = '%s,%s' % (
workspace.name.replace(',','_'),
workspace.name.replace(',','_')
)
for flag in default_options:
if flag not in options:
options.append(flag)
if default_options[flag] is not None:
options.append(default_options[flag])
if not is_paired:
options = remove_pair_options(options)
grace.status('')
full_param = reference.shrimp_command(self.cs, options + reads_parameters)
print >> sys.stderr, 'Running', ' '.join(full_param)
with io.pipe_from(full_param,
stderr=log_file,
cores=cores) as f:
eat(f)
log_file.close()
sam_eater.close()
grace.status('Sort')
#io.execute([
# 'samtools', 'sort', '-n', temp_filename, bam_prefix
#])
sam.sort_bam(temp_filename, bam_prefix, by_name=True, cores=self.cores)
os.unlink(temp_filename)
grace.status('')
