class QualiMapRunner:
QUALIMAP_PATH = '/kb/module/qualimap-bin/qualimap'
def __init__(self, scratch_dir, callback_url, workspace_url, srv_wiz_url):
self.scratch_dir = scratch_dir
self.rau = ReadsAlignmentUtils(callback_url)
self.kbr = KBaseReport(callback_url)
self.dfu = DataFileUtil(callback_url)
self.set_api = SetAPI(srv_wiz_url)
self.ws = Workspace(workspace_url)
self.valid_commands = ['bamqc', 'multi-bamqc']
def run_app(self, params):
self.validate_params(params)
print('Validated Params = ')
pprint(params)
run_info = self.get_run_info(params)
if run_info['mode'] == 'single':
result = self.run_bamqc(params['input_ref'],
run_info['input_info'])
elif run_info['mode'] == 'multi':
result = self.run_multi_sample_qc(params['input_ref'],
run_info['input_info'])
else:
raise ValueError(
'Error in fetching the type to determine run settings.')
if params['create_report']:
result = self.create_report(result, params['output_workspace'])
return result
def create_report(self, result, output_workspace):
qc_result_zip_info = result['qc_result_zip_info']
report_info = self.kbr.create_extended_report({
'message':
'',
'objects_created': [],
'direct_html_link_index':
0,
'html_links': [{
'shock_id': qc_result_zip_info['shock_id'],
'name': qc_result_zip_info['index_html_file_name'],
'label': qc_result_zip_info['name']
}],
'report_object_name':
'qualimap_report' + str(uuid.uuid4()),
'workspace_name':
output_workspace
})
result['report_name'] = report_info['name']
result['report_ref'] = report_info['ref']
return result
def run_bamqc(self, input_ref, input_info):
# download the input and setup a working dir
alignment_info = self.rau.download_alignment({'source_ref': input_ref})
bam_file_path = self.find_my_bam_file(
alignment_info['destination_dir'])
workdir = os.path.join(self.scratch_dir,
'qualimap_' + str(int(time.time() * 10000)))
options = [
'-bam', bam_file_path, '-outdir', workdir, '-outformat', 'html'
]
self.run_cli_command('bamqc', options)
package_info = self.package_output_folder(
workdir, 'QualiMap_report',
'HTML report directory for QualiMap BAM QC', 'qualimapReport.html')
return {
'qc_result_folder_path': workdir,
'qc_result_zip_info': package_info
}
def run_multi_sample_qc(self, input_ref, input_info):
# download the input and setup a working dir
reads_alignment_info = self.get_alignments_from_set(input_ref)
suffix = 'qualimap_' + str(int(time.time() * 10000))
workdir = os.path.join(self.scratch_dir, suffix)
os.makedirs(workdir)
input_file_path = self.create_multi_qualimap_cfg(
reads_alignment_info, workdir)
options = [
'-d', input_file_path, '-r', '-outdir', workdir, '-outformat',
'html'
]
self.run_cli_command('multi-bamqc', options)
package_info = self.package_output_folder(
workdir, 'QualiMap_report',
'HTML report directory for QualiMap Multi-sample BAM QC',
'multisampleBamQcReport.html')
return {
'qc_result_folder_path': workdir,
'qc_result_zip_info': package_info
}
def get_alignments_from_set(self, alignment_set_ref):
set_data = self.set_api.get_reads_alignment_set_v1({
'ref':
alignment_set_ref,
'include_item_info':
1
})
items = set_data['data']['items']
reads_alignment_data = []
for alignment in items:
alignment_info = self.rau.download_alignment(
{'source_ref': alignment['ref']})
bam_file_path = self.find_my_bam_file(
alignment_info['destination_dir'])
label = None
if 'label' in alignment:
label = alignment['label']
reads_alignment_data.append({
'bam_file_path': bam_file_path,
'ref': alignment['ref'],
'label': label,
'info': alignment['info']
})
return reads_alignment_data
def create_multi_qualimap_cfg(self, reads_alignment_info, workdir):
# Group by labels if there is at least one defined
use_labels = False
for alignment in reads_alignment_info:
if alignment['label']:
use_labels = True
break
# write the file
input_file_path = os.path.join(workdir, 'multi_input.txt')
input_file = open(input_file_path, 'w')
name_lookup = {}
for alignment in reads_alignment_info:
name = alignment['info'][1]
if name in name_lookup:
name_lookup[name] += 1
name = name + '_' + str(name_lookup[name])
else:
name_lookup[name] = 1
input_file.write(name + '\t' + alignment['bam_file_path'])
if use_labels:
if alignment['label']:
input_file.write('\t' + alignment['label'])
else:
input_file.write('\tunlabeled')
input_file.write('\n')
input_file.close()
return input_file_path
def get_run_info(self, params):
info = self.get_obj_info(params['input_ref'])
obj_type = self.get_type_from_obj_info(info)
if obj_type in ['KBaseRNASeq.RNASeqAlignment']:
return {'mode': 'single', 'input_info': info}
if obj_type in [
'KBaseRNASeq.RNASeqAlignmentSet', 'KBaseSets.ReadsAlignmentSet'
]:
return {'mode': 'multi', 'input_info': info}
raise ValueError('Object type of input_ref is not valid, was: ' +
str(obj_type))
def validate_params(self, params):
if 'input_ref' not in params:
raise ValueError(
'required parameter field "input_ref" was not set')
create_report = False
if 'create_report' in params:
if int(params['create_report']) == 1:
if 'output_workspace' not in params:
raise ValueError(
'If "create_report" was set, then "output_workspace" is required'
)
if not params['output_workspace']:
raise ValueError(
'If "create_report" was set, then "output_workspace" is required'
)
create_report = True
params['create_report'] = create_report
def run_cli_command(self, command, options, cwd=None):
if command not in self.valid_commands:
raise ValueError('Invalid QualiMap command: ' + str(command))
command = [self.QUALIMAP_PATH, command] + options
print('Running: ' + ' '.join(command))
if not cwd:
cwd = self.scratch_dir
p = subprocess.Popen(command, cwd=cwd, shell=False)
exitCode = p.wait()
if (exitCode == 0):
print('Success, exit code was: ' + str(exitCode))
else:
raise ValueError('Error running command: ' + ' '.join(command) +
'\n' + 'Exit Code: ' + str(exitCode))
def find_my_bam_file(self, dirpath):
bam_path = None
for f in os.listdir(dirpath):
fullpath = os.path.join(dirpath, f)
if os.path.isfile(fullpath) and f.lower().endswith('.bam'):
if bam_path is not None:
raise ValueError(
'Error! Too many BAM files were downloaded for this alignment!'
)
bam_path = fullpath
if bam_path is None:
raise ValueError(
'Error! No BAM files were downloaded for this alignment!')
return bam_path
def package_output_folder(self, folder_path, zip_file_name,
zip_file_description, index_html_file):
''' Simple utility for packaging a folder and saving to shock '''
output = self.dfu.file_to_shock({
'file_path': folder_path,
'make_handle': 0,
'pack': 'zip'
})
return {
'shock_id': output['shock_id'],
'name': zip_file_name,
'description': zip_file_description,
'index_html_file_name': index_html_file
}
def get_type_from_obj_info(self, info):
return info[2].split('-')[0]
def get_obj_info(self, ref):
return self.ws.get_object_info3({'objects': [{
'ref': ref
}]})['infos'][0]
class QualiMapRunner:
QUALIMAP_PATH = '/kb/module/qualimap-bin/qualimap'
JAVA_MEM_DEFAULT_SIZE = '16G'
LARGE_BAM_FILE_SIZE = 20 * 1024 * 1024 * 1024 # 20 GB
TIMEOUT = 72 * 60 * 60 # 72 hours
def _get_file_size(self, file_path):
file_size = os.path.getsize(file_path)
print('File size: {} -- {}'.format(file_size, file_path))
return file_size
def _large_file(self, file_path):
filename, file_extension = os.path.splitext(file_path)
multiplier = 0
if file_extension == '.txt':
total_file_size = 0
with open(file_path, 'r') as f:
for line in f:
bam_file_path = line.split('\t')[1]
total_file_size += self._get_file_size(bam_file_path)
print('Total file size: {}'.format(total_file_size))
multiplier = int(total_file_size) / int(self.LARGE_BAM_FILE_SIZE)
else:
multiplier = int(self._get_file_size(file_path)) / int(
self.LARGE_BAM_FILE_SIZE)
print('setting number of windows multiplier to: {}'.format(multiplier))
return multiplier
def _timeout_handler(self, signum, frame):
print('Signal handler called with signal', signum)
raise ValueError('QualiMap takes too long')
def __init__(self, scratch_dir, callback_url, workspace_url, srv_wiz_url):
self.scratch_dir = scratch_dir
self.rau = ReadsAlignmentUtils(callback_url)
self.kbr = KBaseReport(callback_url)
self.dfu = DataFileUtil(callback_url)
self.gfu = GenomeFileUtil(callback_url)
self.set_api = SetAPI(srv_wiz_url)
self.ws = Workspace(workspace_url)
self.valid_commands = ['bamqc', 'multi-bamqc']
def run_app(self, params):
self.validate_params(params)
print('Validated Params = ')
pprint(params)
run_info = self.get_run_info(params)
if run_info.get('mode') not in ['single', 'multi']:
raise ValueError(
'Error in fetching the type to determine run settings.')
run_error = False
try:
signal.signal(signal.SIGALRM, self._timeout_handler)
signal.alarm(self.TIMEOUT)
if run_info['mode'] == 'single':
result = self.run_bamqc(params['input_ref'],
run_info['input_info'])
elif run_info['mode'] == 'multi':
result = self.run_multi_sample_qc(params['input_ref'],
run_info['input_info'])
signal.alarm(0)
except Exception:
run_error = True
workdir = os.path.join(self.scratch_dir,
'qualimap_' + str(int(time.time() * 10000)))
os.makedirs(workdir)
with open(os.path.join(workdir, 'qualimapReport.html'),
'w') as report:
report.write('<html><body><p></p></body></html>')
package_info = self.package_output_folder(
workdir, 'QualiMap_report',
'EMPTY HTML report directory for QualiMap BAM QC',
'qualimapReport.html')
result = {
'qc_result_folder_path': workdir,
'qc_result_zip_info': package_info,
'shock_id': None
}
error_msg = 'Running QualiMap returned an error:\n{}\n'.format(
traceback.format_exc())
error_msg += 'Generating simple report instead\n'
print(error_msg)
if params['create_report']:
result = self.create_report(result, params['output_workspace'],
run_error, params['input_ref'])
return result
def create_report(self,
result,
output_workspace,
run_error=None,
input_ref=None):
if run_error:
objects_created = []
info = self.get_obj_info(input_ref)
obj_type = self.get_type_from_obj_info(info)
if obj_type in ['KBaseRNASeq.RNASeqAlignment']:
objects_created.append({
'ref': input_ref,
'description': 'Alignment'
})
if obj_type in [
'KBaseRNASeq.RNASeqAlignmentSet',
'KBaseSets.ReadsAlignmentSet'
]:
objects_created.append({
'ref': input_ref,
'description': 'AlignmentSet'
})
reads_alignment_info = self.get_alignments_from_set(input_ref)
for alignment in reads_alignment_info:
alignment_ref = alignment.get('ref')
objects_created.append({
'ref': alignment_ref,
'description': 'Alignment'
})
report_info = self.kbr.create_extended_report({
'message':
' ',
'objects_created':
objects_created,
'report_object_name':
'qualimap_report' + str(uuid.uuid4()),
'workspace_name':
output_workspace
})
result['report_name'] = report_info['name']
result['report_ref'] = report_info['ref']
return result
qc_result_zip_info = result['qc_result_zip_info']
report_info = self.kbr.create_extended_report({
'message':
'',
'objects_created': [],
'direct_html_link_index':
0,
'html_links': [{
'shock_id': qc_result_zip_info['shock_id'],
'name': qc_result_zip_info['index_html_file_name'],
'label': qc_result_zip_info['name']
}],
'report_object_name':
'qualimap_report' + str(uuid.uuid4()),
'workspace_name':
output_workspace
})
result['report_name'] = report_info['name']
result['report_ref'] = report_info['ref']
return result
def get_gtf_file(self, input_ref, set_op=False):
print('Start fetching GFF file from genome')
if set_op:
set_data = self.set_api.get_reads_alignment_set_v1({
'ref':
input_ref,
'include_item_info':
1
})
input_ref = set_data['data']['items'][0]['ref']
obj_data = self.dfu.get_objects({"object_refs":
[input_ref]})['data'][0]['data']
genome_ref = obj_data.get('genome_id')
if not genome_ref:
raise ValueError(
'Alignment is not associated with a Genome object')
result_directory = os.path.join(self.scratch_dir, str(uuid.uuid4()))
os.makedirs(result_directory)
genome_gtf_file = self.gfu.genome_to_gff({
'genome_ref': genome_ref,
'is_gtf': True,
'target_dir': result_directory
})['file_path']
return genome_gtf_file
def run_bamqc(self, input_ref, input_info):
# download the input and setup a working dir
alignment_info = self.rau.download_alignment({'source_ref': input_ref})
bam_file_path = self.find_my_bam_file(
alignment_info['destination_dir'])
try:
gtf_file = self.get_gtf_file(input_ref)
except:
gtf_file = ''
workdir = os.path.join(self.scratch_dir,
'qualimap_' + str(int(time.time() * 10000)))
options = [
'-bam', bam_file_path, '-c', '-outdir', workdir, '-outformat',
'html'
]
if gtf_file:
options += ['-gff', gtf_file]
options.append('--java-mem-size={}'.format(
self.JAVA_MEM_DEFAULT_SIZE)) # always use large mem
multiplier = self._large_file(bam_file_path)
if multiplier:
window_size = multiplier * 400
print('using larger window size: {} and Java memory: {}'.format(
window_size, self.JAVA_MEM_DEFAULT_SIZE))
options.append(
'-nw {}'.format(window_size)) # increase size of windows
self.run_cli_command('bamqc', options)
package_info = self.package_output_folder(
workdir, 'QualiMap_report',
'HTML report directory for QualiMap BAM QC', 'qualimapReport.html')
return {
'qc_result_folder_path': workdir,
'qc_result_zip_info': package_info
}
def run_multi_sample_qc(self, input_ref, input_info):
# download the input and setup a working dir
reads_alignment_info = self.get_alignments_from_set(input_ref)
try:
gtf_file = self.get_gtf_file(input_ref, set_op=True)
except:
gtf_file = ''
suffix = 'qualimap_' + str(int(time.time() * 10000))
workdir = os.path.join(self.scratch_dir, suffix)
os.makedirs(workdir)
input_file_path = self.create_multi_qualimap_cfg(
reads_alignment_info, workdir)
options = [
'-d', input_file_path, '-r', '-c', '-outdir', workdir,
'-outformat', 'html'
]
if gtf_file:
options += ['-gff', gtf_file]
multiplier = self._large_file(input_file_path)
if multiplier:
window_size = multiplier * 400
print('using larger window size: {} and Java memory: {}'.format(
window_size, self.JAVA_MEM_DEFAULT_SIZE))
options.append(
'-nw {}'.format(window_size)) # increase size of windows
options.append('--java-mem-size={}'.format(
self.JAVA_MEM_DEFAULT_SIZE))
self.run_cli_command('multi-bamqc', options)
package_info = self.package_output_folder(
workdir, 'QualiMap_report',
'HTML report directory for QualiMap Multi-sample BAM QC',
'multisampleBamQcReport.html')
return {
'qc_result_folder_path': workdir,
'qc_result_zip_info': package_info
}
def get_alignments_from_set(self, alignment_set_ref):
set_data = self.set_api.get_reads_alignment_set_v1({
'ref':
alignment_set_ref,
'include_item_info':
1
})
items = set_data['data']['items']
reads_alignment_data = []
for alignment in items:
alignment_info = self.rau.download_alignment(
{'source_ref': alignment['ref']})
bam_file_path = self.find_my_bam_file(
alignment_info['destination_dir'])
label = None
if 'label' in alignment:
label = alignment['label']
reads_alignment_data.append({
'bam_file_path': bam_file_path,
'ref': alignment['ref'],
'label': label,
'info': alignment['info']
})
return reads_alignment_data
def create_multi_qualimap_cfg(self, reads_alignment_info, workdir):
# Group by labels if there is at least one defined
use_labels = False
for alignment in reads_alignment_info:
if alignment['label']:
use_labels = True
break
# write the file
input_file_path = os.path.join(workdir, 'multi_input.txt')
input_file = open(input_file_path, 'w')
name_lookup = {}
for alignment in reads_alignment_info:
name = alignment['info'][1]
if name in name_lookup:
name_lookup[name] += 1
name = name + '_' + str(name_lookup[name])
else:
name_lookup[name] = 1
input_file.write(name + '\t' + alignment['bam_file_path'])
if use_labels:
if alignment['label']:
input_file.write('\t' + alignment['label'])
else:
input_file.write('\tunlabeled')
input_file.write('\n')
input_file.close()
return input_file_path
def get_run_info(self, params):
info = self.get_obj_info(params['input_ref'])
obj_type = self.get_type_from_obj_info(info)
if obj_type in ['KBaseRNASeq.RNASeqAlignment']:
return {'mode': 'single', 'input_info': info}
if obj_type in [
'KBaseRNASeq.RNASeqAlignmentSet', 'KBaseSets.ReadsAlignmentSet'
]:
return {'mode': 'multi', 'input_info': info}
raise ValueError('Object type of input_ref is not valid, was: ' +
str(obj_type))
def validate_params(self, params):
if 'input_ref' not in params:
raise ValueError(
'required parameter field "input_ref" was not set')
create_report = False
if 'create_report' in params:
if int(params['create_report']) == 1:
if 'output_workspace' not in params:
raise ValueError(
'If "create_report" was set, then "output_workspace" is required'
)
if not params['output_workspace']:
raise ValueError(
'If "create_report" was set, then "output_workspace" is required'
)
create_report = True
params['create_report'] = create_report
def run_cli_command(self, command, options, cwd=None):
if command not in self.valid_commands:
raise ValueError('Invalid QualiMap command: ' + str(command))
command = [self.QUALIMAP_PATH, command] + options
print('Running: ' + ' '.join(command))
if not cwd:
cwd = self.scratch_dir
p = subprocess.Popen(command, cwd=cwd, shell=False)
exitCode = p.wait()
if (exitCode == 0):
print('Success, exit code was: ' + str(exitCode))
else:
raise ValueError('Error running command: ' + ' '.join(command) +
'\n' + 'Exit Code: ' + str(exitCode))
def find_my_bam_file(self, dirpath):
bam_path = None
for f in os.listdir(dirpath):
fullpath = os.path.join(dirpath, f)
if os.path.isfile(fullpath) and f.lower().endswith('.bam'):
if bam_path is not None:
raise ValueError(
'Error! Too many BAM files were downloaded for this alignment!'
)
bam_path = fullpath
if bam_path is None:
raise ValueError(
'Error! No BAM files were downloaded for this alignment!')
return bam_path
def package_output_folder(self, folder_path, zip_file_name,
zip_file_description, index_html_file):
''' Simple utility for packaging a folder and saving to shock '''
output = self.dfu.file_to_shock({
'file_path': folder_path,
'make_handle': 0,
'pack': 'zip'
})
return {
'shock_id': output['shock_id'],
'name': zip_file_name,
'description': zip_file_description,
'index_html_file_name': index_html_file
}
def get_type_from_obj_info(self, info):
return info[2].split('-')[0]
def get_obj_info(self, ref):
return self.ws.get_object_info3({'objects': [{
'ref': ref
}]})['infos'][0]