def create_analysis_dir(project,
top_dir=None,
merge_replicates=False,
keep_names=False,
dry_run=False):
"""Create and populate analysis directory for an IlluminaProject
Creates a new directory and populates either with links to FASTQ
files, or with 'merged' FASTQ files created by concatenating
multiple FASTQs for each sample (which can happen for multiplexed
runs where samples are split across multiple lanes).
Project directory names are made up of the project name and then
the experiment type, or just the project name if experiment type
is not set.
Arguments:
project : populated IlluminaProject object
top_dir : parent directory to create analysis subdirectory
under. Defaults to cwd if not explicitly specified
merge_replicates: if True then creates a single FASTQ file for
each sample by merging multiple FASTQs together
keep_names: if True then links to FASTQ files will have the same
names as the original files; by default links use the
shortest unique name
dry_run : if True then report what would be done but don't
actually perform any action
Returns:
Name of the project directory.
"""
project_dir = os.path.join(top_dir,project.full_name)
print "Creating analysis directory for project '%s'..." % project.full_name
# Check for & create directory
if os.path.exists(project_dir):
print "-> %s already exists" % project_dir
else:
print "Making analysis directory for %s" % project.name
if not dry_run:
bcf_utils.mkdir(project_dir,mode=0775)
# Make an empty ScriptCode directory
scriptcode_dir = os.path.join(project_dir,"ScriptCode")
if os.path.exists(scriptcode_dir):
print "'ScriptCode' directory %s already exists" % scriptcode_dir
else:
print "Making 'ScriptCode' directory for %s" % project.name
if not dry_run:
bcf_utils.mkdir(scriptcode_dir,mode=0775)
# Check for & create links to fastq files
if not merge_replicates:
for sample in project.samples:
fastq_names = IlluminaData.get_unique_fastq_names(sample.fastq)
for fastq in sample.fastq:
fastq_file = os.path.join(sample.dirn,fastq)
if keep_names:
fastq_ln = os.path.join(project_dir,fastq)
else:
fastq_ln = os.path.join(project_dir,fastq_names[fastq])
if os.path.exists(fastq_ln):
logging.error("Failed to link to %s: %s already exists" %
(fastq_file,os.path.basename(fastq_ln)))
else:
print "Linking to %s" % fastq
if not dry_run:
bcf_utils.mklink(fastq_file,fastq_ln,relative=True)
else:
# Merge files for replicates within each sample
for sample in project.samples:
replicates = {}
# Gather replicates to be merged
for fastq in sample.fastq:
fastq_data = IlluminaData.IlluminaFastq(fastq)
name = "%s_%s_R%d" % (fastq_data.sample_name,
fastq_data.barcode_sequence,
fastq_data.read_number)
if name not in replicates:
replicates[name] = []
replicates[name].append(os.path.join(sample.dirn,fastq))
# Sort into order
replicates[name].sort()
# Report detected replicates
print "Sample %s" % sample.name
for name in replicates:
print "\tReplicate '%s'" % name
for fastq in replicates[name]:
print "\t\t%s" % fastq
# Do the merge
for name in replicates:
merged_fastq = os.path.join(project_dir,name+'.fastq')
bcf_utils.concatenate_fastq_files(merged_fastq,replicates[name])
# Return directory name
return project_dir
"if required)")
p.add_option_group(deprecated_options)
# Process command line
options,args = p.parse_args()
if len(args) != 1:
p.error("input is a single SampleSheet.csv file")
if options.miseq:
logging.warning("--miseq option no longer necessary; MiSEQ-style sample sheets "
"are now converted automatically")
# Get input sample sheet file
samplesheet = args[0]
if not os.path.isfile(samplesheet):
logging.error("sample sheet '%s': not found" % samplesheet)
sys.exit(1)
# Read in the data as CSV
data = IlluminaData.get_casava_sample_sheet(samplesheet)
# Remove lanes
if options.lanes is not None:
lanes = parse_lane_expression(options.lanes)
print "Keeping lanes %s, removing the rest" % ','.join([str(x) for x in lanes])
new_data = IlluminaData.CasavaSampleSheet()
for line in data:
if line['Lane'] in lanes:
print "Keeping %s" % line
new_data.append(tabdata="%s" % line)
data = new_data
# Update the SampleID and SampleProject fields
for sample_id in options.sample_id:
lanes,name = parse_name_expression(sample_id)
for line in data:
if line['Lane'] in lanes:
"'NY_ChIP-seq'. Use multiple --expt=... to set the types for different "
"projects")
p.add_option("--keep-names",action="store_true",dest="keep_names",default=False,
help="preserve the full names of the source fastq files when creating links")
p.add_option("--merge-replicates",action="store_true",dest="merge_replicates",default=False,
help="create merged fastq files for each set of replicates detected")
# Parse command line
options,args = p.parse_args()
# Get data directory name
if len(args) != 1:
p.error("expected one argument (location of Illumina analysis dir)")
illumina_analysis_dir = os.path.abspath(args[0])
# Populate Illumina data object
illumina_data = IlluminaData.IlluminaData(illumina_analysis_dir,
unaligned_dir=options.unaligned_dir)
# Assign experiment types
for expt in options.expt_type:
name,type_ = expt.split(':')
illumina_data.get_project(name).expt_type = type_
# Create and populate per-project directory structure
for project in illumina_data.projects:
create_analysis_dir(project,
top_dir=illumina_analysis_dir,
merge_replicates=options.merge_replicates,
keep_names=options.keep_names,
dry_run=options.dry_run)
"if required)")
p.add_option_group(deprecated_options)
# Process command line
options,args = p.parse_args()
if len(args) != 1:
p.error("input is a single SampleSheet.csv file")
if options.miseq:
logging.warning("--miseq option no longer necessary; MiSEQ-style sample sheets "
"are now converted automatically")
# Get input sample sheet file
samplesheet = args[0]
if not os.path.isfile(samplesheet):
logging.error("sample sheet '%s': not found" % samplesheet)
sys.exit(1)
# Read in the data as CSV
data = IlluminaData.get_casava_sample_sheet(samplesheet)
# Update the SampleID and SampleProject fields
for sample_id in options.sample_id:
lanes,name = parse_name_expression(sample_id)
for line in data:
if line['Lane'] in lanes:
print "Setting SampleID for lane %d: '%s'" % (line['Lane'],name)
line['SampleID'] = name
# Update the SampleProject field
for sample_project in options.sample_project:
lanes,name = parse_name_expression(sample_project)
for line in data:
if line['Lane'] in lanes:
print "Setting SampleProject for lane %d: '%s'" % (line['Lane'],name)
line['SampleProject'] = name
# Fix spaces
p.add_option('--ignore-warnings',action="store_true",dest="ignore_warnings",default=False,
help="ignore warnings about spaces and duplicated sampleID/sampleProject "
"combinations when writing new samplesheet.csv file")
# Process command line
options,args = p.parse_args()
if len(args) != 1:
p.error("input is a single SampleSheet.csv file")
# Get input sample sheet file
samplesheet = args[0]
if not os.path.isfile(samplesheet):
logging.error("sample sheet '%s': not found" % samplesheet)
sys.exit(1)
# Read in the data as CSV
if options.miseq:
# Input sample sheet is from MiSEQ
data = IlluminaData.convert_miseq_samplesheet_to_casava(samplesheet)
else:
# Standard CASAVA sample sheet
data = IlluminaData.CasavaSampleSheet(samplesheet)
# Update the SampleID and SampleProject fields
for sample_id in options.sample_id:
lanes,name = parse_name_expression(sample_id)
for line in data:
if line['Lane'] in lanes:
print "Setting SampleID for lane %d: '%s'" % (line['Lane'],name)
line['SampleID'] = name
# Update the SampleProject field
for sample_project in options.sample_project:
lanes,name = parse_name_expression(sample_project)
for line in data:
if line['Lane'] in lanes:
def demultiplex_fastq(fastq_file, barcodes, nmismatches):
"""Perform demultiplexing of a FASTQ file
Demultiplex reads in a FASTQ file given information about a set of
barcode/index sequences.
Produces a file for each barcode, plus another for 'unbinned'
reads.
Arguments:
fastq_file: FASTQ file to be demultiplexed (can be gzipped)
barcodes: list of barcode sequences to use for demultiplexing
nmismatches: maxiumum number of mismatched bases allowed when
testing whether barcode sequences match
Returns:
No return value
"""
# Start
print "Processing %s" % fastq_file
info = IlluminaData.IlluminaFastq(fastq_file)
# Set up output files
output_files = {}
# Weed out barcodes that aren't associated with this lane
local_barcodes = []
for barcode in barcodes:
if barcode['lane'] != info.lane_number:
continue
local_barcodes.append(barcode)
output_file_name = "%s_%s_L%03d_R%d_%03d.fastq" % (
barcode['name'], barcode['index'], info.lane_number,
info.read_number, info.set_number)
print "\t%s\t%s" % (barcode['index'], output_file_name)
if os.path.exists(output_file_name):
print "\t%s: already exists,exiting" % output_file_name
sys.exit(1)
output_files[barcode['index']] = open(output_file_name, 'w')
# Check if there's anything to do
if len(local_barcodes) == 0:
return
# Also make a file for unbinned reads
unbinned_file_name = "unbinned_L%03d_R%d_%03d.fastq" % (
info.lane_number, info.read_number, info.set_number)
if os.path.exists(unbinned_file_name):
print "\t%s: already exists,exiting" % unbinned_file_name
sys.exit(1)
output_files['unbinned'] = open(unbinned_file_name, 'w')
# Process reads
nreads = 0
for read in FASTQFile.FastqIterator(fastq_file):
nreads += 1
matched_read = False
this_barcode = read.seqid.index_sequence
for barcode in local_barcodes:
if barcode['matcher'].match(this_barcode, nmismatches):
##print "Matched %s against %s" % (this_barcode,barcodes[barcode]['name'])
output_files[barcode['index']].write(str(read) + '\n')
matched_read = True
break
# Put in unbinned if no match
if not matched_read:
output_files['unbinned'].write(str(read) + '\n')
##if nreads > 100: break
# Close files
for barcode in local_barcodes:
output_files[barcode['index']].close()
print "\tMatched %d reads for %s" % (nreads, os.path.basename(fastq_file))
# Set up barcode data
barcodes = []
for barcode_info in options.barcode_info:
name, barcode, lane = barcode_info.split(':')
print "Assigning barcode '%s' in lane %s to %s" % (barcode, lane, name)
barcodes.append({
'name': name,
'index': barcode,
'matcher': BarcodeMatcher(barcode),
'lane': int(lane)
})
# Read from sample sheet (if supplied)
if options.sample_sheet is not None:
print "Reading data from sample sheet %s" % options.sample_sheet
sample_sheet = IlluminaData.CasavaSampleSheet(options.sample_sheet)
for line in sample_sheet:
name = line['SampleID']
barcode = line['Index'].rstrip('N').rstrip('-').rstrip('N')
lane = line['Lane']
print "Assigning barcode '%s' in lane %s to %s" % (barcode, lane,
name)
barcodes.append({
'name': name,
'index': barcode,
'matcher': BarcodeMatcher(barcode),
'lane': int(lane)
})
if len(barcodes) < 1:
p.error("need at least one --barcode and/or --samplesheet assignment")
else:
n_fastqs = len(sample.fastq)
if n_fastqs == 1:
print "\t%s" % sample.name
else:
print "\t%s (%d fastqs)" % (sample.name,n_fastqs)
# Print fastq names
fastqs = sample.fastq_subset(read_number=1) + \
sample.fastq_subset(read_number=2)
for fastq in fastqs:
print "\t\t%s" % fastq
# Report the names of the samples in each project
if options.report:
for project in illumina_data.projects:
print "%s" % IlluminaData.describe_project(project)
# Report statistics for fastq files
if options.stats:
# Print number of reads for each file, and file size
for sample in project.samples:
for fastq in sample.fastq:
fq = os.path.join(sample.dirn,fastq)
nreads = FASTQFile.nreads(fq)
fsize = os.path.getsize(fq)
print "%s\t%s\t%d" % (fastq,
bcf_utils.format_file_size(fsize),
nreads)
print ""
# Summary: short report suitable for logging file
if options.summary: