def alignment_post_processing(BASECALLER_RESULTS, ALIGNMENT_RESULTS, flows,
mark_duplicates, force_alignstats):
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"),
'r')
datasets_basecaller = json.load(f)
f.close()
except:
printtime("ERROR: problem parsing %s" %
os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 400
input_prefix_list = []
for dataset in datasets_basecaller["datasets"]:
if not os.path.exists(
os.path.join(BASECALLER_RESULTS, dataset['basecaller_bam'])):
continue
printtime("Barcode processing, rename")
src = os.path.join(ALIGNMENT_RESULTS,
dataset['file_prefix'] + '.alignment.summary')
if os.path.exists(src):
input_prefix_list.append(
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix'] + '.'))
#terrible hack to make aggregate_alignment happy
X_name = 'nomatch'
read_group = dataset['read_groups'][0]
if 'barcode_name' in datasets_basecaller['read_groups'][
read_group]:
X_name = datasets_basecaller['read_groups'][read_group][
'barcode_name']
dst = os.path.join(ALIGNMENT_RESULTS,
'alignment_%s.summary' % X_name)
try:
os.symlink(os.path.relpath(src, os.path.dirname(dst)), dst)
except:
printtime("ERROR: Unable to symlink '%s' to '%s'" % (src, dst))
# Special legacy post-processing.
# Generate merged rawlib.bam on barcoded runs
composite_bam_filename = os.path.join(ALIGNMENT_RESULTS, 'rawlib.bam')
if not os.path.exists(composite_bam_filename):
bam_file_list = []
for dataset in datasets_basecaller["datasets"]:
bam_name = os.path.join(
ALIGNMENT_RESULTS,
os.path.basename(dataset['file_prefix']) + '.bam')
if os.path.exists(bam_name):
bam_file_list.append(bam_name)
blockprocessing.merge_bam_files(bam_file_list, composite_bam_filename,
composite_bam_filename + '.bai',
mark_duplicates)
force_alignstats = True
if force_alignstats:
## Generate data for error plot for barcoded run from composite bam
printtime("Call alignStats to generate raw accuracy")
try:
cmd = "alignStats"
cmd += " -n 12"
cmd += " --alignSummaryFile alignStats_err.txt"
cmd += " --alignSummaryJsonFile alignStats_err.json"
cmd += " --alignSummaryMinLen 1"
#cmd += " --alignSummaryMaxLen %s" % str(int(graph_max_x))
cmd += " --alignSummaryMaxLen %s" % str(int(400))
cmd += " --alignSummaryLenStep 1"
cmd += " --alignSummaryMaxErr 10"
cmd += " --infile %s" % composite_bam_filename
cmd = cmd + " --outputDir %s" % ALIGNMENT_RESULTS
printtime("DEBUG: Calling '%s'" % cmd)
os.system(cmd)
except:
printtime("alignStats failed")
mergeAlignStatsResults(input_prefix_list, ALIGNMENT_RESULTS + "/")
try:
base_error_plot.generate_base_error_plot(
os.path.join(ALIGNMENT_RESULTS, 'alignStats_err.json'),
os.path.join(ALIGNMENT_RESULTS, 'base_error_plot.png'),
int(graph_max_x))
ionstats_plots.alignment_rate_plot(
os.path.join(ALIGNMENT_RESULTS, 'alignStats_err.json'),
os.path.join(BASECALLER_RESULTS, 'ionstats_basecaller.json'),
os.path.join(ALIGNMENT_RESULTS, 'alignment_rate_plot.png'),
int(graph_max_x))
# Create aligned histogram plot
# Create AQ20 plot
printtime("Base error plot has been created successfully")
except:
printtime("ERROR: Failed to generate base error plot")
traceback.print_exc()
# Generate alignment_barcode_summary.csv
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment("./", barcodelist_path)
# These graphs are likely obsolete
makeAlignGraphs()
def merge_alignment_stats(dirs, BASECALLER_RESULTS, ALIGNMENT_RESULTS, flows):
datasets_json = {}
try:
f = open(os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"),
'r')
datasets_json = json.load(f)
f.close()
except:
printtime("ERROR: problem parsing %s" %
os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"))
traceback.print_exc()
return
for dataset in datasets_json['datasets']:
# What needs merging:
# - alignment.summary
# - alignTable.txt
# Some time in the future:
# - alignStats_err.json
# Merge alignStats metrics
try:
input_prefix_list = [
os.path.join(dir, ALIGNMENT_RESULTS,
dataset['file_prefix'] + '.') for dir in dirs
]
input_prefix_list = [
prefix for prefix in input_prefix_list
if os.path.exists(prefix + 'alignment.summary')
]
composite_prefix = os.path.join(ALIGNMENT_RESULTS,
dataset['file_prefix'] + '.')
if input_prefix_list:
mergeAlignStatsResults(input_prefix_list, composite_prefix)
else:
printtime("Nothing to merge: " + dataset['file_prefix'])
except:
printtime("ERROR: merging %s stats unsuccessful" %
(dataset['file_prefix'] + '.bam'))
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"),
'r')
datasets_basecaller = json.load(f)
f.close()
except:
printtime("ERROR: problem parsing %s" %
os.path.join(BASECALLER_RESULTS, "datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 400
input_prefix_list = []
for dataset in datasets_basecaller["datasets"]:
printtime("Barcode processing, rename")
src = os.path.join(ALIGNMENT_RESULTS,
dataset['file_prefix'] + '.alignment.summary')
if os.path.exists(src):
input_prefix_list.append(
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix'] + '.'))
#terrible hack to make aggregate_alignment happy
X_name = 'nomatch'
read_group = dataset['read_groups'][0]
if 'barcode_name' in datasets_basecaller['read_groups'][
read_group]:
X_name = datasets_basecaller['read_groups'][read_group][
'barcode_name']
dst = os.path.join(ALIGNMENT_RESULTS,
'alignment_%s.summary' % X_name)
try:
os.symlink(os.path.relpath(src, os.path.dirname(dst)), dst)
except:
printtime("ERROR: Unable to symlink '%s' to '%s'" % (src, dst))
# Merge alignStats_err.json right here!
merged_align_stats = {}
align_stats_num_bases = 400
for dir in dirs:
current_align_stats = {}
try:
f = open(
os.path.join(dir, ALIGNMENT_RESULTS, 'alignStats_err.json'),
'r')
current_align_stats = json.load(f)
f.close()
except:
printtime(
"Merge alignStats_err.json: skipping %s" %
os.path.join(dir, ALIGNMENT_RESULTS, 'alignStats_err.json'))
continue
if not merged_align_stats:
merged_align_stats = current_align_stats
align_stats_num_bases = len(
merged_align_stats.get("read_length", []))
continue
for idx in range(align_stats_num_bases):
merged_align_stats['nread'][idx] += current_align_stats['nread'][
idx]
merged_align_stats['unaligned'][idx] += current_align_stats[
'unaligned'][idx]
merged_align_stats['filtered'][idx] += current_align_stats[
'filtered'][idx]
merged_align_stats['clipped'][idx] += current_align_stats[
'clipped'][idx]
merged_align_stats['aligned'][idx] += current_align_stats[
'aligned'][idx]
merged_align_stats['n_err_at_position'][
idx] += current_align_stats['n_err_at_position'][idx]
merged_align_stats['cum_aligned'][idx] += current_align_stats[
'cum_aligned'][idx]
merged_align_stats['cum_err_at_position'][
idx] += current_align_stats['cum_err_at_position'][idx]
merged_align_stats['accuracy_total_bases'] += current_align_stats[
'accuracy_total_bases']
merged_align_stats['accuracy_total_errors'] += current_align_stats[
'accuracy_total_errors']
merged_align_stats['total_mapped_target_bases'] += current_align_stats[
'total_mapped_target_bases']
merged_align_stats['total_mapped_reads'] += current_align_stats[
'total_mapped_reads']
try:
f = open(os.path.join(ALIGNMENT_RESULTS, 'alignStats_err.json'), "w")
json.dump(merged_align_stats, f, indent=4)
f.close()
except:
printtime("ERROR; Failed to write merged alignStats_err.json")
traceback.print_exc()
mergeAlignStatsResults(input_prefix_list, ALIGNMENT_RESULTS + "/")
try:
base_error_plot.generate_base_error_plot(
os.path.join(ALIGNMENT_RESULTS, 'alignStats_err.json'),
os.path.join(ALIGNMENT_RESULTS, 'base_error_plot.png'),
int(graph_max_x))
ionstats_plots.alignment_rate_plot(
os.path.join(ALIGNMENT_RESULTS, 'alignStats_err.json'),
os.path.join(BASECALLER_RESULTS, 'ionstats_basecaller.json'),
os.path.join(ALIGNMENT_RESULTS, 'alignment_rate_plot.png'),
int(graph_max_x))
printtime("Base error plot has been created successfully")
except:
printtime("ERROR: Failed to generate base error plot")
traceback.print_exc()
# Generate alignment_barcode_summary.csv
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment("./", barcodelist_path)
def alignment_post_processing(
libraryName,
BASECALLER_RESULTS,
ALIGNMENT_RESULTS,
flows,
mark_duplicates):
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"),'r')
datasets_basecaller = json.load(f)
f.close()
except:
printtime("ERROR: problem parsing %s" % os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 800
alignment_file_list = []
for dataset in datasets_basecaller["datasets"]:
if not os.path.exists(os.path.join(BASECALLER_RESULTS, dataset['basecaller_bam'])):
continue
ionstats.generate_ionstats_alignment(
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.bam'),
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.ionstats_alignment.json'),
graph_max_x)
ionstats2alignstats(libraryName,
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.ionstats_alignment.json'),
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.alignment.summary'))
alignment_file_list.append(os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.ionstats_alignment.json'))
# In Progress: merge ionstats alignment results
ionstats.reduce_stats(alignment_file_list,os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'))
ionstats2alignstats(libraryName,
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
os.path.join(ALIGNMENT_RESULTS,'alignment.summary'))
# Special legacy post-processing.
# Generate merged rawlib.bam on barcoded runs
composite_bam_filename = os.path.join(ALIGNMENT_RESULTS,'rawlib.bam')
if not os.path.exists(composite_bam_filename):
bam_file_list = []
for dataset in datasets_basecaller["datasets"]:
bam_name = os.path.join(ALIGNMENT_RESULTS,os.path.basename(dataset['file_prefix'])+'.bam')
if os.path.exists(bam_name):
bam_file_list.append(bam_name)
blockprocessing.merge_bam_files(bam_file_list,composite_bam_filename,composite_bam_filename+'.bai',mark_duplicates)
# Generate alignment_barcode_summary.csv
#TODO: use datasets_basecaller.json + *.ionstats_alignment.json instead of barcodeList.txt and alignment_*.summary
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment ("./",barcodelist_path)
# These graphs are likely obsolete
#makeAlignGraphs()
# In Progress: Use ionstats alignment results to generate plots
ionstats_plots.alignment_rate_plot2(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'alignment_rate_plot.png', graph_max_x)
ionstats_plots.base_error_plot(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'base_error_plot.png', graph_max_x)
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q10.png', 'AQ10', 'red')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q17.png', 'AQ17', 'yellow')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q20.png', 'AQ20', 'green')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q47.png', 'AQ47', 'purple')
def merge_alignment_stats(dirs, BASECALLER_RESULTS, ALIGNMENT_RESULTS, flows):
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"),'r')
datasets_basecaller = json.load(f)
f.close()
except:
printtime("ERROR: problem parsing %s" % os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 800
########################################################
# Merge ionstats_alignment.json
# First across blocks, then across barcoded
########################################################
try:
composite_filename_list = []
for dataset in datasets_basecaller["datasets"]:
composite_filename = os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.ionstats_alignment.json')
barcode_filename_list = [os.path.join(dir,ALIGNMENT_RESULTS,dataset['file_prefix']+'.ionstats_alignment.json') for dir in dirs]
barcode_filename_list = [filename for filename in barcode_filename_list if os.path.exists(filename)]
ionstats.reduce_stats(barcode_filename_list,composite_filename)
if os.path.exists(composite_filename):
composite_filename_list.append(composite_filename)
ionstats.reduce_stats(composite_filename_list,os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'))
except:
printtime("ERROR: Failed to merge ionstats_alignment.json")
traceback.print_exc()
# Use ionstats alignment results to generate plots
ionstats_plots.alignment_rate_plot2(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'alignment_rate_plot.png', graph_max_x)
ionstats_plots.base_error_plot(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'base_error_plot.png', graph_max_x)
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q10.png', 'AQ10', 'red')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q17.png', 'AQ17', 'yellow')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q20.png', 'AQ20', 'green')
ionstats_plots.old_aq_length_histogram(
os.path.join(ALIGNMENT_RESULTS,'ionstats_alignment.json'),
'Filtered_Alignments_Q47.png', 'AQ47', 'purple')
# Generate alignment_barcode_summary.csv
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment ("./",barcodelist_path)
def merge_alignment_stats(dirs, BASECALLER_RESULTS, ALIGNMENT_RESULTS, flows):
datasets_json = {}
try:
f = open(os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"),'r')
datasets_json = json.load(f);
f.close()
except:
printtime("ERROR: problem parsing %s" % os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"))
traceback.print_exc()
return
for dataset in datasets_json['datasets']:
# What needs merging:
# - alignment.summary
# - alignTable.txt
# Some time in the future:
# - alignStats_err.json
# Merge alignStats metrics
try:
input_prefix_list = [os.path.join(dir,ALIGNMENT_RESULTS, dataset['file_prefix']+'.') for dir in dirs]
input_prefix_list = [prefix for prefix in input_prefix_list if os.path.exists(prefix+'alignment.summary')]
composite_prefix = os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.')
if input_prefix_list:
mergeAlignStatsResults(input_prefix_list,composite_prefix)
else:
printtime("Nothing to merge: "+dataset['file_prefix'])
except:
printtime("ERROR: merging %s stats unsuccessful" % (dataset['file_prefix']+'.bam'))
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"),'r')
datasets_basecaller = json.load(f);
f.close()
except:
printtime("ERROR: problem parsing %s" % os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 400
input_prefix_list = []
for dataset in datasets_basecaller["datasets"]:
printtime("Barcode processing, rename")
src = os.path.join(ALIGNMENT_RESULTS,dataset['file_prefix']+'.alignment.summary')
if os.path.exists(src):
input_prefix_list.append(os.path.join(ALIGNMENT_RESULTS,dataset['file_prefix']+'.'))
#terrible hack to make aggregate_alignment happy
X_name = 'nomatch'
read_group = dataset['read_groups'][0]
if 'barcode_name' in datasets_basecaller['read_groups'][read_group]:
X_name = datasets_basecaller['read_groups'][read_group]['barcode_name']
dst = os.path.join(ALIGNMENT_RESULTS, 'alignment_%s.summary' % X_name)
try:
os.symlink(os.path.relpath(src,os.path.dirname(dst)),dst)
except:
printtime("ERROR: Unable to symlink '%s' to '%s'" % (src, dst))
# Merge alignStats_err.json right here!
merged_align_stats = {}
align_stats_num_bases = 400
for dir in dirs:
current_align_stats = {}
try:
f = open(os.path.join(dir,ALIGNMENT_RESULTS,'alignStats_err.json'),'r')
current_align_stats = json.load(f);
f.close()
except:
printtime("Merge alignStats_err.json: skipping %s" % os.path.join(dir,ALIGNMENT_RESULTS,'alignStats_err.json'))
continue
if not merged_align_stats:
merged_align_stats = current_align_stats
align_stats_num_bases = len(merged_align_stats.get("read_length",[]))
continue
for idx in range(align_stats_num_bases):
merged_align_stats['nread'][idx] += current_align_stats['nread'][idx]
merged_align_stats['unaligned'][idx] += current_align_stats['unaligned'][idx]
merged_align_stats['filtered'][idx] += current_align_stats['filtered'][idx]
merged_align_stats['clipped'][idx] += current_align_stats['clipped'][idx]
merged_align_stats['aligned'][idx] += current_align_stats['aligned'][idx]
merged_align_stats['n_err_at_position'][idx] += current_align_stats['n_err_at_position'][idx]
merged_align_stats['cum_aligned'][idx] += current_align_stats['cum_aligned'][idx]
merged_align_stats['cum_err_at_position'][idx] += current_align_stats['cum_err_at_position'][idx]
merged_align_stats['accuracy_total_bases'] += current_align_stats['accuracy_total_bases']
merged_align_stats['accuracy_total_errors'] += current_align_stats['accuracy_total_errors']
merged_align_stats['total_mapped_target_bases'] += current_align_stats['total_mapped_target_bases']
merged_align_stats['total_mapped_reads'] += current_align_stats['total_mapped_reads']
try:
f = open(os.path.join(ALIGNMENT_RESULTS,'alignStats_err.json'),"w")
json.dump(merged_align_stats, f, indent=4)
f.close()
except:
printtime("ERROR; Failed to write merged alignStats_err.json")
traceback.print_exc()
mergeAlignStatsResults(input_prefix_list,ALIGNMENT_RESULTS+"/")
try:
base_error_plot.generate_base_error_plot(
os.path.join(ALIGNMENT_RESULTS,'alignStats_err.json'),
os.path.join(ALIGNMENT_RESULTS,'base_error_plot.png'),int(graph_max_x))
base_error_plot.generate_alignment_rate_plot(
os.path.join(ALIGNMENT_RESULTS,'alignStats_err.json'),
os.path.join(BASECALLER_RESULTS,'readLen.txt'),
os.path.join(ALIGNMENT_RESULTS,'alignment_rate_plot.png'),int(graph_max_x))
printtime("Base error plot has been created successfully")
except:
printtime("ERROR: Failed to generate base error plot")
traceback.print_exc()
# Generate alignment_barcode_summary.csv
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment ("./",barcodelist_path)
def alignment_post_processing(
BASECALLER_RESULTS,
ALIGNMENT_RESULTS,
flows,
mark_duplicates,
force_alignstats):
datasets_basecaller = {}
try:
f = open(os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"),'r')
datasets_basecaller = json.load(f);
f.close()
except:
printtime("ERROR: problem parsing %s" % os.path.join(BASECALLER_RESULTS,"datasets_basecaller.json"))
traceback.print_exc()
return
try:
graph_max_x = int(50 * math.ceil(0.014 * int(flows)))
except:
graph_max_x = 400
input_prefix_list = []
for dataset in datasets_basecaller["datasets"]:
if not os.path.exists(os.path.join(BASECALLER_RESULTS, dataset['basecaller_bam'])):
continue
printtime("Barcode processing, rename")
src = os.path.join(ALIGNMENT_RESULTS,dataset['file_prefix']+'.alignment.summary')
if os.path.exists(src):
input_prefix_list.append(os.path.join(ALIGNMENT_RESULTS,dataset['file_prefix']+'.'))
#terrible hack to make aggregate_alignment happy
X_name = 'nomatch'
read_group = dataset['read_groups'][0]
if 'barcode_name' in datasets_basecaller['read_groups'][read_group]:
X_name = datasets_basecaller['read_groups'][read_group]['barcode_name']
dst = os.path.join(ALIGNMENT_RESULTS, 'alignment_%s.summary' % X_name)
try:
os.symlink(os.path.relpath(src,os.path.dirname(dst)),dst)
except:
printtime("ERROR: Unable to symlink '%s' to '%s'" % (src, dst))
printtime("Creating legacy name links")
if dataset.has_key('legacy_prefix'):
link_src = [
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.bam'),
os.path.join(ALIGNMENT_RESULTS, dataset['file_prefix']+'.bam.bai')]
link_dst = [
os.path.join(ALIGNMENT_RESULTS, os.path.basename(dataset['legacy_prefix'])+'.bam'),
os.path.join(ALIGNMENT_RESULTS, os.path.basename(dataset['legacy_prefix'])+'.bam.bai')]
for (src,dst) in zip(link_src,link_dst):
try:
os.symlink(os.path.relpath(src,os.path.dirname(dst)),dst)
except:
printtime("ERROR: Unable to symlink '%s' to '%s'" % (src, dst))
# Special legacy post-processing.
# Generate merged rawlib.basecaller.bam and rawlib.sff on barcoded runs
composite_bam_filename = os.path.join(ALIGNMENT_RESULTS,'rawlib.bam')
if not os.path.exists(composite_bam_filename):
bam_file_list = []
for dataset in datasets_basecaller["datasets"]:
bam_name = os.path.join(ALIGNMENT_RESULTS,os.path.basename(dataset['file_prefix'])+'.bam')
if os.path.exists(bam_name):
bam_file_list.append(bam_name)
blockprocessing.merge_bam_files(bam_file_list,composite_bam_filename,composite_bam_filename+'.bai',mark_duplicates)
force_alignstats = True
if force_alignstats:
## Generate data for error plot for barcoded run from composite bam
printtime("Call alignStats to generate raw accuracy")
try:
cmd = "alignStats"
cmd += " -n 12"
cmd += " --alignSummaryFile alignStats_err.txt"
cmd += " --alignSummaryJsonFile alignStats_err.json"
cmd += " --alignSummaryMinLen 1"
#cmd += " --alignSummaryMaxLen %s" % str(int(graph_max_x))
cmd += " --alignSummaryMaxLen %s" % str(int(400))
cmd += " --alignSummaryLenStep 1"
cmd += " --alignSummaryMaxErr 10"
cmd += " --infile %s" % composite_bam_filename
cmd = cmd + " --outputDir %s" % ALIGNMENT_RESULTS
printtime("DEBUG: Calling '%s'" % cmd)
os.system(cmd)
except:
printtime("alignStats failed")
mergeAlignStatsResults(input_prefix_list,ALIGNMENT_RESULTS+"/")
try:
base_error_plot.generate_base_error_plot(
os.path.join(ALIGNMENT_RESULTS,'alignStats_err.json'),
os.path.join(ALIGNMENT_RESULTS,'base_error_plot.png'),int(graph_max_x))
base_error_plot.generate_alignment_rate_plot(
os.path.join(ALIGNMENT_RESULTS,'alignStats_err.json'),
os.path.join(BASECALLER_RESULTS,'readLen.txt'),
os.path.join(ALIGNMENT_RESULTS,'alignment_rate_plot.png'),int(graph_max_x))
# Create aligned histogram plot
# Create AQ20 plot
printtime("Base error plot has been created successfully")
except:
printtime("ERROR: Failed to generate base error plot")
traceback.print_exc()
# Generate alignment_barcode_summary.csv
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if os.path.exists(barcodelist_path):
printtime("Barcode processing, aggregate")
aggregate_alignment ("./",barcodelist_path)
# These graphs are likely obsolete
makeAlignGraphs()
def align_barcodes(
sammeta,
libsff_path,
align_full,
sam_parsed,
bidirectional,
libraryName,
DIR_BC_FILES,
flows,
aligner_opts_extra,
mark_duplicates,
ALIGNMENT_RESULTS,
outBaseName=''
):
printtime("Renaming non-barcoded alignment results to 'comprehensive'")
files = [ 'alignment.summary',
'alignmentQC_out.txt',
'alignTable.txt',
]
for fname in files:
if os.path.exists(fname):
try:
#if os.path.exists(fname):
# os.rename(fname, fname + ".comprehensive")
shutil.copyfile(fname, fname + ".comprehensive")
except:
printtime('ERROR copying %s' % fname)
traceback.print_exc()
printtime("STARTING BARCODE ALIGNMENTS")
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
printtime('ERROR: barcodeList.txt not found')
barcodeList = parse_bcfile(barcodelist_path)
align_full = True
(head,tail) = os.path.split(libsff_path)
for bcid in (x['id_str'] for x in barcodeList):
sffName = os.path.join(DIR_BC_FILES,"%s_%s" % (bcid, tail))
print "sffName: "+sffName
if os.path.exists(sffName):
printtime("Barcode processing for '%s': %s" % (bcid, sffName))
else:
printtime("No barcode SFF file found for '%s': %s" % (bcid, sffName))
continue
align_full_chip(
sammeta,
sffName,
align_full,
1,
True,
sam_parsed,
bidirectional,
libraryName,
flows,
aligner_opts_extra,
mark_duplicates,
ALIGNMENT_RESULTS,
outBaseName)
#rename each output file based on barcode found in fastq filename
#but ignore the comprehensive fastq output files
printtime("Barcode processing, rename")
if os.path.exists('alignment.summary'):
try:
fname='alignment_%s.summary' % bcid
os.rename('alignment.summary', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignmentQC_out_%s.txt' % bcid
os.rename('alignmentQC_out.txt', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignTable_%s.txt' % bcid
os.rename('alignTable.txt', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
except:
printtime('error renaming')
traceback.print_exc()
#rename comprehensive results back to default names
for fname in files:
if os.path.exists(fname + '.comprehensive'):
# os.rename(fname + '.comprehensive', fname)
try:
shutil.copyfile(fname + '.comprehensive', fname)
except:
printtime('ERROR copying %s' % fname + '.comprehensive')
traceback.print_exc()
printtime("Barcode processing, aggreagate")
aggregate_alignment ("./",barcodelist_path)
def align_full_chip_core(libsff, libKey, tfKey, floworder, fastqName, align_full, graph_max_x, do_barcode, make_align_graphs, sam_parsed, DIR_BC_FILES, env, outputdir):
#collect all the meta data for the SAM file
SAM_META = {}
# id - this hash comes from the fastq file
try:
#read the first line of the fastq file
fastqFile = open(fastqName,'r')
id_hash = fastqFile.readline()
fastqFile.close()
#now just pull out the hash
id_hash = id_hash[1:6]
SAM_META['ID'] = id_hash
except IOError:
printtime("Could not read fastq file. The ID for the SAM file could not be found.")
# sm - name for reads - project name
SAM_META['SM'] = env['project']
# lb - library name
SAM_META['LB'] = env['libraryName']
# pu - the platform unit
SAM_META['PU'] = "PGM/" + env['chipType'].replace('"',"")
SAM_META['PL'] = "IONTORRENT"
#TODO: do not assume localhost. Find the name of the masternode
try:
#this will get the exp data from the database
exp_json = json.loads(env['exp_json'])
# ds - the "notes", only the alphanumeric and space characters.
SAM_META['DS'] = ''.join(ch for ch in exp_json['notes'] if ch.isalnum() or ch == " ")
# dt - the run date
exp_log_json = json.loads(exp_json['log'])
iso_exp_time = exp_log_json['start_time']
#convert to ISO time
iso_exp_time = dateutil.parser.parse(iso_exp_time)
SAM_META['DT'] = iso_exp_time.isoformat()
#the site name should be here, also remove spaces
site_name = env['site_name']
site_name = ''.join(ch for ch in site_name if ch.isalnum() )
SAM_META['CN'] = site_name
env['flows'] = exp_json['flows']
except:
printtime("There was an error getting the site name, because the Torrent Browser could not be contacted")
traceback.print_exc()
#Now build the SAM meta data arg string
aligner_opts_rg= '--aligner-opts-rg "'
aligner_opts_extra = ''
if sam_parsed:
aligner_opts_extra += ' -p 1'
if env['aligner_opts_extra']:
print ' found extra alignment options: "%s"' % env['aligner_opts_extra']
aligner_opts_extra = ' --aligner-opts-extra "'
aligner_opts_extra += env['aligner_opts_extra'] + '"'
first = True
for key, value in SAM_META.items():
if value:
sam_arg = r'-R \"'
end = r'\"'
sam_arg = sam_arg + key + ":" + value + end
if first:
aligner_opts_rg = aligner_opts_rg + sam_arg
first = False
else:
aligner_opts_rg = aligner_opts_rg + " " + sam_arg
#add the trailing quote
aligner_opts_rg = aligner_opts_rg + '"'
if 0 < graph_max_x:
# establish the read-length histogram range by using the simple rule: 0.6 * num-flows
flowsUsed = 0
try:
flowsUsed = int(env['flows'])
except:
flowsUsed = 400
graph_max_x = 100 * math.trunc((0.6 * flowsUsed + 99)/100.0)
if graph_max_x < 400:
graph_max_x = 400
#-----------------------------------
# DEFAULT SINGLE SFF/FASTQ BEHAVIOR - (Runs for barcoded runs too)
#-----------------------------------
if (align_full):
#If a full align is forced add a '--align-all-reads' flag
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % libsff
com += " --genome %s" % env["libraryName"]
com += " --max-plot-read-len %s" % graph_max_x
com += " --align-all-reads"
com += " %s %s" % (aligner_opts_rg,aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
else:
# Add -p 1 to enable default.sam file generation
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % libsff
com += " --genome %s" % env["libraryName"]
com += " --max-plot-read-len %s" % graph_max_x
com += " %s %s" % (aligner_opts_rg,aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
try:
printtime("Alignment QC command line:\n%s" % com)
retcode = subprocess.call(com, shell=True)
if retcode != 0:
printtime("alignmentQC failed, return code: %d" % retcode)
alignError = open("alignment.error", "w")
alignError.write('alignmentQC returned with error code: ')
alignError.write(str(retcode))
alignError.close()
except OSError:
printtime('Alignment Failed to start')
alignError = open("alignment.error", "w")
alignError.write(str(traceback.format_exc()))
alignError.close()
traceback.print_exc()
if make_align_graphs:
makeAlignGraphs()
#--------------------------------------------
# BARCODE HANDLING BEHAVIOR (Multiple FASTQ)
#--------------------------------------------
if env['barcodeId'] and True == do_barcode:
printtime("Renaming non-barcoded alignment results to 'comprehensive'")
files = [ 'alignment.summary',
'alignmentQC_out.txt',
'alignTable.txt',
]
for fname in files:
try:
if os.path.exists(fname):
os.rename(fname, fname + ".comprehensive")
except:
printtime('error renaming')
traceback.print_exc()
# Only make the graphs from the alignment of comprehensive fastq file
if make_align_graphs:
makeAlignGraphs()
printtime("STARTING BARCODE ALIGNMENTS")
if not os.path.exists(DIR_BC_FILES):
os.mkdir(DIR_BC_FILES)
barcodeList = parse_bcfile('barcodeList.txt')
align_full = True
for bcid in (x['id_str'] for x in barcodeList):
sffName = "%s_%s_%s.sff" % (bcid, env['expName'], env['resultsName'])
if not os.path.exists(sffName):
printtime("No barcode SFF file found for '%s'" % bcid)
continue
if (align_full):
printtime("Align All Reads")
#If a full align is forced add a '--align-all-reads' flag
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % sffName
com += " --genome %s" % env["libraryName"]
com += " --max-plot-read-len %s" % graph_max_x
com += " --align-all-reads"
com += " %s %s" % (aligner_opts_rg, aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
else:
printtime("Align Subset of Reads")
# Add -p 1 to enable default.sam file generation
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % sffName
com += " --genome %s" % env["libraryName"]
com += " --max-plot-read-len %s" % graph_max_x
com += " %s %s" % (aligner_opts_rg, aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
try:
printtime("Alignment QC command line:\n%s" % com)
retcode = subprocess.call(com, shell=True)
if retcode != 0:
printtime("alignmentQC failed, return code: %d" % retcode)
alignError = open("alignment.error", "a")
alignError.write(com)
alignError.write(': \nalignmentQC returned with error code: ')
alignError.write(str(retcode))
alignError.close()
except OSError:
printtime('Alignment Failed to start')
alignError = open("alignment.error", "a")
alignError.write(str(traceback.format_exc()))
alignError.close()
traceback.print_exc()
#rename each output file based on barcode found in fastq filename
#but ignore the comprehensive fastq output files
if os.path.exists('alignment.summary'):
try:
fname='alignment_%s.summary' % bcid
os.rename('alignment.summary', fname)
os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignmentQC_out_%s.txt' % bcid
os.rename('alignmentQC_out.txt', fname)
os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignTable_%s.txt' % bcid
os.rename('alignTable.txt', fname)
os.rename(fname,os.path.join(DIR_BC_FILES,fname))
#move fastq, sff, bam, bai files
extlist = ['fastq','sff','bam','bam.bai']
for ext in extlist:
bcfile = "%s_%s_%s.%s" % (bcid,env['expName'], env['resultsName'],ext)
if os.path.isfile(bcfile):
os.rename(bcfile,os.path.join(DIR_BC_FILES,bcfile))
except:
printtime('error renaming')
traceback.print_exc()
#rename comprehensive results back to default names
files = [ 'alignment.summary',
'alignmentQC_out.txt',
'alignTable.txt',
]
for fname in files:
if os.path.exists(fname + '.comprehensive'):
os.rename(fname + '.comprehensive', fname)
aggregate_alignment (DIR_BC_FILES,'barcodeList.txt')
def align_full_chip(
SAM_META,
libsff_path,
align_full,
graph_max_x,
do_barcode,
make_align_graphs,
sam_parsed,
bidirectional,
DIR_BC_FILES,
libraryName,
flows,
barcodeId,
opts_extra,
outputdir):
printtime("sam_parsed is %s" % sam_parsed)
#Now build the SAM meta data arg string
aligner_opts_rg= '--aligner-opts-rg "'
aligner_opts_extra = ''
additional_aligner_opts = ''
if sam_parsed:
additional_aligner_opts += ' -p 1'
if bidirectional:
additional_aligner_opts += ' --bidirectional'
if opts_extra:
print ' found extra alignment options: "%s"' % opts_extra
aligner_opts_extra = ' --aligner-opts-extra "'
aligner_opts_extra += opts_extra + '"'
first = True
for key, value in SAM_META.items():
if value:
sam_arg = r'-R \"'
end = r'\"'
sam_arg = sam_arg + key + ":" + value + end
if first:
aligner_opts_rg = aligner_opts_rg + sam_arg
first = False
else:
aligner_opts_rg = aligner_opts_rg + " " + sam_arg
#add the trailing quote
aligner_opts_rg = aligner_opts_rg + '"'
if 0 < graph_max_x:
# establish the read-length histogram range by using the simple rule: 0.6 * num-flows
flowsUsed = 0
try:
flowsUsed = int(flows)
except:
flowsUsed = 400
graph_max_x = 100 * math.trunc((0.6 * flowsUsed + 99)/100.0)
if graph_max_x < 400:
graph_max_x = 400
#-----------------------------------
# DEFAULT SINGLE SFF/FASTQ BEHAVIOR - (Runs for barcoded runs too)
#-----------------------------------
if (align_full):
#If a full align is forced add a '--align-all-reads' flag
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % libsff_path
com += " --genome %s" % libraryName
com += " --max-plot-read-len %s" % graph_max_x
com += " --align-all-reads"
com += " %s" % (additional_aligner_opts)
com += " %s %s" % (aligner_opts_rg,aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
else:
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % libsff_path
com += " --genome %s" % libraryName
com += " --max-plot-read-len %s" % graph_max_x
com += " %s" % (additional_aligner_opts)
com += " %s %s" % (aligner_opts_rg,aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
try:
printtime("Alignment QC command line:\n%s" % com)
retcode = subprocess.call(com, shell=True)
blockprocessing.add_status("alignmentQC.pl", retcode)
if retcode != 0:
printtime("alignmentQC failed, return code: %d" % retcode)
alignError = open("alignment.error", "w")
alignError.write('alignmentQC returned with error code: ')
alignError.write(str(retcode))
alignError.close()
except OSError:
printtime('Alignment Failed to start')
alignError = open("alignment.error", "w")
alignError.write(str(traceback.format_exc()))
alignError.close()
traceback.print_exc()
if make_align_graphs:
makeAlignGraphs()
#--------------------------------------------
# BARCODE HANDLING BEHAVIOR (Multiple FASTQ)
#--------------------------------------------
if barcodeId and do_barcode:
printtime("Renaming non-barcoded alignment results to 'comprehensive'")
files = [ 'alignment.summary',
'alignmentQC_out.txt',
'alignTable.txt',
]
for fname in files:
try:
#if os.path.exists(fname):
# os.rename(fname, fname + ".comprehensive")
shutil.copyfile(fname, fname + ".comprehensive")
except:
printtime('ERROR copying %s' % fname)
traceback.print_exc()
printtime("STARTING BARCODE ALIGNMENTS")
barcodelist_path = 'barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../barcodeList.txt'
if not os.path.exists(barcodelist_path):
barcodelist_path = '../../barcodeList.txt'
if not os.path.exists(barcodelist_path):
printtime('ERROR: barcodeList.txt not found')
barcodeList = parse_bcfile(barcodelist_path)
align_full = True
top_dir = os.getcwd()
try:
os.chdir(DIR_BC_FILES)
printtime('DEBUG changing to %s for barcodes alignment' % DIR_BC_FILES)
except:
printtime('ERROR missing %s folder' % DIR_BC_FILES)
for bcid in (x['id_str'] for x in barcodeList):
(head,tail) = os.path.split(libsff_path)
sffName = os.path.join(head,"%s_%s" % (bcid, tail))
if os.path.exists(sffName):
printtime("Barcode processing for '%s': %s" % (bcid, sffName))
else:
printtime("No barcode SFF file found for '%s': %s" % (bcid, sffName))
continue
if (align_full):
printtime("Align All Reads")
#If a full align is forced add a '--align-all-reads' flag
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % sffName
com += " --genome %s" % libraryName
com += " --max-plot-read-len %s" % graph_max_x
com += " --align-all-reads"
com += " %s" % (additional_aligner_opts)
com += " %s %s" % (aligner_opts_rg, aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
else:
printtime("Align Subset of Reads")
com = "alignmentQC.pl"
com += " --logfile %s" % os.path.join(outputdir,"alignmentQC_out.txt")
com += " --output-dir %s" % outputdir
com += " --input %s" % sffName
com += " --genome %s" % libraryName
com += " --max-plot-read-len %s" % graph_max_x
com += " %s" % (additional_aligner_opts)
com += " %s %s" % (aligner_opts_rg, aligner_opts_extra)
com += " >> ReportLog.html 2>&1"
try:
printtime("Alignment QC command line:\n%s" % com)
retcode = subprocess.call(com, shell=True)
blockprocessing.add_status("alignmentQC.pl", retcode)
if retcode != 0:
printtime("alignmentQC failed, return code: %d" % retcode)
alignError = open("alignment.error", "a")
alignError.write(com)
alignError.write(': \nalignmentQC returned with error code: ')
alignError.write(str(retcode))
alignError.close()
except:
printtime('ERROR: Alignment Failed to start')
alignError = open("alignment.error", "a")
alignError.write(str(traceback.format_exc()))
alignError.close()
traceback.print_exc()
#rename each output file based on barcode found in fastq filename
#but ignore the comprehensive fastq output files
if os.path.exists('alignment.summary'):
try:
fname='alignment_%s.summary' % bcid
os.rename('alignment.summary', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignmentQC_out_%s.txt' % bcid
os.rename('alignmentQC_out.txt', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
fname='alignTable_%s.txt' % bcid
os.rename('alignTable.txt', fname)
# os.rename(fname,os.path.join(DIR_BC_FILES,fname))
except:
printtime('error renaming')
traceback.print_exc()
os.chdir(top_dir)
#rename comprehensive results back to default names
for fname in files:
#if os.path.exists(fname + '.comprehensive'):
# os.rename(fname + '.comprehensive', fname)
try:
shutil.copyfile(fname + '.comprehensive', fname)
except:
printtime('ERROR copying %s' % fname + '.comprehensive')
traceback.print_exc()
aggregate_alignment (DIR_BC_FILES,barcodelist_path)