def get_git(name, description, source, url, commit, checksum, destination,
preparation, checker):
'''
Get a dependency from git
'''
if _os.path.realpath(_os.path.curdir) != destination:
try:
_os.chdir(destination)
except OSError:
_os.makedirs(destination)
_os.chdir(destination)
try:
# clear any previous verions
_shutil.rmtree(url.split('/')[-1].replace('.git', ''))
except OSError:
pass
git_server = url.replace('https://', '').replace('http://',
'').split('/')[0]
print('Downloading {} via git from {} . . .'.format(name, git_server))
_subprocess.call(['git', 'clone', url])
_os.chdir(url.split('/')[-1].replace('.git', ''))
_subprocess.call(['git', 'checkout', commit])
# if repo uses git submodules, those will be set to the correct revisions for this commit
# else will do nothing
_subprocess.call(['git', 'submodule', 'update', '--init'])
working_dir = _os.path.sep.join(
[destination, url.split('/')[-1].replace('.git', '')])
if preparation is not None:
for do_this in preparation:
if isinstance(do_this['arguments'], dict):
do_this['function'](**do_this['arguments'])
else:
do_this['function'](*do_this['arguments'])
# restore position in path if a prepare changed it
if working_dir != _os.path.realpath(_os.path.curdir):
_os.chdir(working_dir)
_os.chdir(_os.path.pardir)
_os.chdir(_os.path.pardir)
def get_git(name, description, source, url, commit, checksum, destination, preparation, checker):
'''
Get a dependency from git
'''
if _os.path.realpath(_os.path.curdir) != destination:
try:
_os.chdir(destination)
except OSError:
_os.makedirs(destination)
_os.chdir(destination)
try:
# clear any previous verions
_shutil.rmtree(url.split('/')[-1].replace('.git',''))
except OSError:
pass
git_server = url.replace('https://','').replace('http://','').split('/')[0]
print('Downloading {} via git from {} . . .'.format(name, git_server))
_subprocess.call(['git', 'clone', url])
_os.chdir(url.split('/')[-1].replace('.git',''))
_subprocess.call(['git', 'checkout', commit])
# if repo uses git submodules, those will be set to the correct revisions for this commit
# else will do nothing
_subprocess.call(['git', 'submodule', 'update', '--init'])
working_dir = _os.path.sep.join([destination,url.split('/')[-1].replace('.git','')])
if preparation is not None:
for do_this in preparation:
if isinstance(do_this['arguments'], dict):
do_this['function'](**do_this['arguments'])
else:
do_this['function'](*do_this['arguments'])
# restore position in path if a prepare changed it
if working_dir != _os.path.realpath(_os.path.curdir):
_os.chdir(working_dir)
_os.chdir(_os.path.pardir)
_os.chdir(_os.path.pardir)
def getFromENA(self, run_acc_list,
ftp_server_url = 'ftp.sra.ebi.ac.uk',
local_reads_path = ['reads']):
'''
Given a list of 'run' accession numbers for paired end short read analyses,
download the read files from the European Nucleotide Archive.
If using a mirror server, supply an alternative for 'ftp_server_url'.
'local_reads_path' can be a path string or list or folder names.
'''
if isinstance(local_reads_path, list):
local_reads_path = _os.path.sep.join(local_reads_path)
if not _os.path.exists(local_reads_path):
_os.makedirs(local_reads_path)
print('Logging in to %s' % ftp_server_url)
ftp = _FTP(ftp_server_url)
# anonymous login
print(ftp.login())
def check_connection(ftp):
try:
print('FTP: %s' % ftp.voidcmd("NOOP"))
# http://docs.python.org/2/library/ftplib.html
return(True)
except IOError as e:
print('Seems to be a problem with the connection to FTP server:')
print('I/O error({0}): {1}'.format(e.errno, e.strerror) )
return(False)
def calc_checksum(filepath):
hasher = _md5()
handle = open(filepath, 'rb')
buff = handle.read(65536)
while len(buff) > 0:
hasher.update(buff)
buff = handle.read(65536)
return(hasher.hexdigest())
downloaded_read_files = {}
start_time = _time.time()
failed = []
for cnum,run_acc in enumerate(run_acc_list):
query_url_base = 'http://www.ebi.ac.uk/ena/data/warehouse/search?query='
success = False
tries = 0
max_tries = 5
while not success:
rest_req = '"run_accession=%s"&result=read_run&fields=fastq_ftp,fastq_md5&display=report' % run_acc
print('Sending query to ENA:\n%s' % rest_req)
result = _urllib2.urlopen(query_url_base + rest_req).read()
print('ENA accession numbers query result:\n%s' % result)
if result.count('ERR') == 7:
success = True
else:
print('Query result from ENA was unexpected on attempt %s of %s' % (tries, max_tries))
_time.sleep(0.5)
tries += 1
if tries == max_tries:
print('Attempt %s failed. Try again later and if problem persists, report bug.' % tries)
failed += [run_acc]
break
#_sys.exit(1)
if not success:
continue
md5s = result.split('\n')[-2].split('\t')[-1][:-1].split(';')
ENA_paths = result.split('\n')[-2].split('\t')[-2][:-1].split(';')
ENA_reads_pair_paths = {}
ENA_reads_pair_paths[1] = ENA_paths[0].replace(ftp_server_url, '')
ENA_reads_pair_paths[2] = ENA_paths[1].replace(ftp_server_url, '')
local_reads_pair_paths = {}
local_reads_pair_paths[1] = local_reads_path + \
_os.path.sep + \
ENA_reads_pair_paths[1].split('/')[-1]
local_reads_pair_paths[2] = local_reads_path + \
_os.path.sep + \
ENA_reads_pair_paths[2].split('/')[-1]
downloaded_read_files[run_acc] = {}
for f in (1,2):
# ensure connection is still open
while not check_connection(ftp):
_sleep(0.5)
print('Attempting to re-establish connection . . .')
ftp = _FTP(ftp_server_url)
# anonymous login
print(ftp.login())
pass
expected_checksum = md5s[f - 1]
exists = _os.path.exists(local_reads_pair_paths[f])
if exists:
print('File %s for %s exists locally: %s' % (f, run_acc, local_reads_pair_paths[f]))
actual_checksum = calc_checksum(local_reads_pair_paths[f])
if actual_checksum == expected_checksum:
print('File checksum matches: %s. Skipping download' % (expected_checksum))
downloaded_read_files[run_acc][f] = local_reads_pair_paths[f]
continue
else:
print('Checksum mismatch')
print('Downloading via %s: %s' % (ftp_server_url, ENA_reads_pair_paths[f]))
res = ftp.retrbinary('RETR %s' % ENA_reads_pair_paths[f],
open(local_reads_pair_paths[f], 'wb').write)
print('FTP: %s' % res)
print('Calculating checksum . . .')
actual_checksum = calc_checksum(local_reads_pair_paths[f])
if actual_checksum == expected_checksum:
print('File checksum matches: %s.' % (expected_checksum))
downloaded_read_files[run_acc][f] = local_reads_pair_paths[f]
else:
print('Checksum mismatch for: %s')
if len(run_acc_list) > 1:
# report durations, time left etc
_report_time(start_time, cnum, len(run_acc_list))
if len(failed) > 0:
print('WARNING: some accession numbers did not return a result from ENA')
print('Try searching http://www.ebi.ac.uk/ena in a web-browser for:')
print(', '.join(failed))
self.read_files = downloaded_read_files
def getFromENA(self,
run_acc_list,
ftp_server_url='ftp.sra.ebi.ac.uk',
local_reads_path=['reads']):
'''
Given a list of 'run' accession numbers for paired end short read analyses,
download the read files from the European Nucleotide Archive.
If using a mirror server, supply an alternative for 'ftp_server_url'.
'local_reads_path' can be a path string or list or folder names.
'''
if isinstance(local_reads_path, list):
local_reads_path = _os.path.sep.join(local_reads_path)
if not _os.path.exists(local_reads_path):
_os.makedirs(local_reads_path)
print('Logging in to %s' % ftp_server_url)
ftp = _FTP(ftp_server_url)
# anonymous login
print(ftp.login())
def check_connection(ftp):
try:
print('FTP: %s' % ftp.voidcmd("NOOP"))
# http://docs.python.org/2/library/ftplib.html
return (True)
except IOError as e:
print(
'Seems to be a problem with the connection to FTP server:')
print('I/O error({0}): {1}'.format(e.errno, e.strerror))
return (False)
def calc_checksum(filepath):
hasher = _md5()
handle = open(filepath, 'rb')
buff = handle.read(65536)
while len(buff) > 0:
hasher.update(buff)
buff = handle.read(65536)
return (hasher.hexdigest())
downloaded_read_files = {}
start_time = _time.time()
failed = []
for cnum, run_acc in enumerate(run_acc_list):
query_url_base = 'http://www.ebi.ac.uk/ena/data/warehouse/search?query='
success = False
tries = 0
max_tries = 5
while not success:
rest_req = '"run_accession=%s"&result=read_run&fields=fastq_ftp,fastq_md5&display=report' % run_acc
print('Sending query to ENA:\n%s' % rest_req)
result = _urllib2.urlopen(query_url_base + rest_req).read()
print('ENA accession numbers query result:\n%s' % result)
if result.count('ERR') == 7:
success = True
else:
print(
'Query result from ENA was unexpected on attempt %s of %s'
% (tries, max_tries))
_time.sleep(0.5)
tries += 1
if tries == max_tries:
print(
'Attempt %s failed. Try again later and if problem persists, report bug.'
% tries)
failed += [run_acc]
break
#_sys.exit(1)
if not success:
continue
md5s = result.split('\n')[-2].split('\t')[-1][:-1].split(';')
ENA_paths = result.split('\n')[-2].split('\t')[-2][:-1].split(';')
ENA_reads_pair_paths = {}
ENA_reads_pair_paths[1] = ENA_paths[0].replace(ftp_server_url, '')
ENA_reads_pair_paths[2] = ENA_paths[1].replace(ftp_server_url, '')
local_reads_pair_paths = {}
local_reads_pair_paths[1] = local_reads_path + \
_os.path.sep + \
ENA_reads_pair_paths[1].split('/')[-1]
local_reads_pair_paths[2] = local_reads_path + \
_os.path.sep + \
ENA_reads_pair_paths[2].split('/')[-1]
downloaded_read_files[run_acc] = {}
for f in (1, 2):
# ensure connection is still open
while not check_connection(ftp):
_sleep(0.5)
print('Attempting to re-establish connection . . .')
ftp = _FTP(ftp_server_url)
# anonymous login
print(ftp.login())
pass
expected_checksum = md5s[f - 1]
exists = _os.path.exists(local_reads_pair_paths[f])
if exists:
print('File %s for %s exists locally: %s' %
(f, run_acc, local_reads_pair_paths[f]))
actual_checksum = calc_checksum(local_reads_pair_paths[f])
if actual_checksum == expected_checksum:
print('File checksum matches: %s. Skipping download' %
(expected_checksum))
downloaded_read_files[run_acc][
f] = local_reads_pair_paths[f]
continue
else:
print('Checksum mismatch')
print('Downloading via %s: %s' %
(ftp_server_url, ENA_reads_pair_paths[f]))
res = ftp.retrbinary(
'RETR %s' % ENA_reads_pair_paths[f],
open(local_reads_pair_paths[f], 'wb').write)
print('FTP: %s' % res)
print('Calculating checksum . . .')
actual_checksum = calc_checksum(local_reads_pair_paths[f])
if actual_checksum == expected_checksum:
print('File checksum matches: %s.' % (expected_checksum))
downloaded_read_files[run_acc][f] = local_reads_pair_paths[
f]
else:
print('Checksum mismatch for: %s')
if len(run_acc_list) > 1:
# report durations, time left etc
_report_time(start_time, cnum, len(run_acc_list))
if len(failed) > 0:
print(
'WARNING: some accession numbers did not return a result from ENA'
)
print(
'Try searching http://www.ebi.ac.uk/ena in a web-browser for:')
print(', '.join(failed))
self.read_files = downloaded_read_files
def SPAdes(self,
exe=[],
output_folder=['assemblies', 'SPAdes'],
mem_num_gigs=8,
max_cpus=-1,
single_assembly=False,
careful=True,
only_assembler=False):
'''
de novo assembly of short reads using SPAdes
By default, the provided short reads in dictionary: self.paths_to_reads
will be assembled separately, unless single_assembly set to True in
which case each set of paired read fastq files will be used in a
single assembly.
http://spades.bioinf.spbau.ru/release3.6.1/manual.html
relevent inputs:
-o <output_dir> Specify the output directory. Required option.
--sc required for MDA (single-cell) data.
--only-error-correction
--only-assembler
--careful reduce the number of mismatches and short indels. Run MismatchCorrector – a post processing tool. Recommended.
--continue from the specified output folder starting from the last available check-point
--restart-from <check_point>
ec start from error correction
as restart assembly module from the first iteration
k<int> restart from the iteration with specified k values, e.g. k55
mc restart mismatch correction
--pe1-12 <file_name> interlaced forward and reverse paired-end reads.
--pe1-1 <file_name> File with forward reads.
--pe1-2 <file_name> File with reverse reads.
--pe1-s <file_name> File with unpaired reads . . use --pe2-... for next library
--threads <int>
--memory <int> max memory in Gb
-k <int,int,...> Comma-separated list of odd ascending k-mers
If --sc is set the default value are 21,33,55, for multicell data sets it is auto
--cov-cutoff <float> positive float value, or 'auto', or 'off'. Default value is 'off'
'''
assert isinstance(
output_folder,
list), 'Provide output folder as list of folders forming path'
base_output_path = _os.path.sep.join(output_folder)
if not _os.path.exists(base_output_path):
_os.makedirs(base_output_path)
# max threads is slightly different to cpus
# . . can probably use more
max_processes = _decide_max_processes(max_cpus)
# if an exe is not provided, use that stored in Dependencies
if len(exe):
use_exe = _os.path.sep.join(exe)
else:
from baga import Dependencies
use_exe = _get_exe_path('spades')
def run_SPAdes(cmd):
proc = _subprocess.Popen(cmd,
stdout=_subprocess.PIPE,
stderr=_subprocess.PIPE)
# allow for failed SPAdes runs (possibly caused by small fastq files) <== but also check they were actually built properly
try:
stdout_value, stderr_value = proc.communicate()
checkthese = []
getline = False
for line in stdout_value.split('\n'):
if 'Warnings saved to' in line:
getline = False
if getline:
l = line.rstrip()
if len(l):
checkthese += [l]
if 'SPAdes pipeline finished WITH WARNINGS!' in line:
getline = True
if len(checkthese):
print('SPAdes completed with warnings:\n{}\n'.format(
'\n'.join(checkthese)))
else:
print('SPAdes completed without warnings')
# with open('___SPAdes_{}_good_{}.log'.format(cnum, thetime), 'w') as fout:
# fout.write(stdout_value)
path2contigs = _os.path.sep.join(
[this_output_path, 'contigs.fasta'])
except _subprocess.CalledProcessError as e:
print('SPAdes probably did not complete: error returned ({})'.
format(proc.returncode))
print('Error: {}'.format(e))
print(
'Writing some info relevent to SPAdes crash to ___SPAdes_{}_bad_{}.log'
.format(cnum, thetime))
with open('___SPAdes_{}_bad_{}.log'.format(cnum, thetime),
'w') as fout:
fout.write(dir(proc))
fout.write('\n' + str(e.returncode) + '\n')
fout.write(
_os.path.sep.join([this_output_path, 'contigs.fasta']))
path2contigs = None
return (path2contigs)
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
contigs = {}
if single_assembly:
print(
'Combining reads aligned at multiple regions into single assembly'
)
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
for cnum, (pairname, files) in enumerate(self.read_files.items()):
# allow use of tuples or dicts by converting dicts to lists
if isinstance(files, dict):
use_files = []
for k, v in sorted(files.items()):
use_files += [v]
else:
use_files = files
cmd += ['--pe{}-1'.format(cnum + 1), use_files[0]]
cmd += ['--pe{}-2'.format(cnum + 1), use_files[1]]
try:
# use unpaired reads if available
cmd += ['--pe{}-s'.format(cnum + 1), use_files[2]]
except IndexError:
pass
try:
# add a second library if provided
if isinstance(self.read_files2[pairname], dict):
# if a dict supplied, make it a list
use_files2 = []
for k, v in sorted(self.read_files2[pairname].items()):
use_files2 += [v]
else:
use_files2 = self.read_files2[pairname]
cmd += ['--pe{}-1'.format(cnum + 2), use_files2[0]]
cmd += ['--pe{}-2'.format(cnum + 2), use_files2[1]]
try:
cmd += ['--pe{}-s'.format(cnum + 2), use_files2[2]]
except IndexError:
pass
except AttributeError:
pass
## this isn't very flexible:
# retain <sample>__<genome> from pairname:
# pairname == <sample>__<genome>_<start>-<end>+<padding>
# and replace with multiregion
folder = '{}__{}_{}'.format(
pairname.split('__')[0],
pairname.split('__')[1].split('_')[0], 'multi_region')
this_output_path = _os.path.sep.join(output_folder + [folder])
if not _os.path.exists(this_output_path):
_os.makedirs(this_output_path)
cmd += ['-o', this_output_path]
cmd += ['--threads', str(max_processes)]
cmd += ['--memory', str(mem_num_gigs)]
if only_assembler:
cmd += ['--only-assembler']
if careful:
cmd += ['--careful']
thetime = _time.asctime(_time.localtime(_time.time()))
print('about to launch SPAdes . . . at {}'.format(thetime))
print(' '.join(cmd))
contigs['multi_region'] = run_SPAdes(cmd)
else:
start_time = _time.time()
# prepare commandline and launch each SPAdes assembly
contigs = {}
for cnum, (pairname,
files) in enumerate(sorted(self.read_files.items())):
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
# allow use of tuples or dicts by converting dicts to lists
if isinstance(files, dict):
use_files = []
for k, v in sorted(files.items()):
use_files += [v]
else:
use_files = files
cmd += ['--pe1-1', use_files[0]]
cmd += ['--pe1-2', use_files[1]]
try:
# use unpaired reads if available
cmd += ['--pe1-s', use_files[2]]
except IndexError:
pass
try:
# add a second library if provided
if isinstance(self.read_files2[pairname], dict):
# if a dict supplied, make it a list
use_files2 = []
for k, v in sorted(self.read_files2[pairname].items()):
use_files2 += [v]
else:
use_files2 = self.read_files2[pairname]
cmd += ['--pe2-1', use_files2[0]]
cmd += ['--pe2-2', use_files2[1]]
try:
cmd += ['--pe2-s', use_files2[2]]
except IndexError:
pass
except AttributeError:
pass
this_output_path = _os.path.sep.join(output_folder +
[pairname])
if not _os.path.exists(this_output_path):
_os.makedirs(this_output_path)
cmd += ['-o', this_output_path]
cmd += ['--threads', str(max_processes)]
cmd += ['--memory', str(mem_num_gigs)]
if only_assembler:
cmd += ['--only-assembler']
if careful:
cmd += ['--careful']
thetime = _time.asctime(_time.localtime(_time.time()))
print('about to launch SPAdes . . . at {}'.format(thetime))
print(' '.join(cmd))
contigs[pairname] = run_SPAdes(cmd)
if len(self.read_files) > 1:
# report durations, time left etc
_report_time(start_time, cnum, len(self.read_files))
self.paths_to_contigs = contigs
def get_download(name, description, source, url, commit, checksum, destination,
preparation, checker):
'''
Download and unpack a dependancy
'''
##
initialdir = _os.path.abspath(_os.curdir)
try:
_os.chdir(destination)
except OSError:
_os.makedirs(destination)
_os.chdir(destination)
if checksum:
hasher_algorithm = checksum.split('=')[0]
if hasher_algorithm == 'md5':
hasher = _hashlib.md5()
elif hasher_algorithm == 'sha1':
hasher = _hashlib.sha1()
elif hasher_algorithm == 'sha224':
hasher = _hashlib.sha224()
elif hasher_algorithm == 'sha256':
hasher = _hashlib.sha256()
elif hasher_algorithm == 'sha384':
hasher = _hashlib.sha384()
elif hasher_algorithm == 'sha512':
hasher = _hashlib.sha512()
else:
print("{} checksums not implemented in Python's hashlib!".format(
hasher_algorithm))
print('Downloading: %s' % url)
req = _urllib2.urlopen(url)
CHUNK = 16 * 1024 * 16
data = _cStringIO.StringIO()
c = 0
for chunk in iter(lambda: req.read(CHUNK), ''):
c += CHUNK
print("{:,} bytes".format(c))
data.write(chunk)
print('Download complete . . .')
data.seek(0)
if checksum:
buff = data.read(65536)
while len(buff) > 0:
hasher.update(buff)
buff = data.read(65536)
e = '. . . checksum fail!'
assert hasher.hexdigest() == checksum.split('=')[1], e
print('. . . checksum passed!')
data.seek(0)
if url[-6:] == 'tar.gz':
archive = _tarfile.open(mode="r:gz", fileobj=data)
elif url[-7:] == 'tar.bz2':
archive = _tarfile.open(mode="r:bz2", fileobj=data)
elif url[-4:] == '.zip':
archive = _zipfile.ZipFile(data)
if destination == 'local_packages':
# extract as a pypi python package
release = url.split('/')[-1][:-7]
print('Extracting {} to {}'.format(
release, _os.path.sep.join([destination, name])))
c = 0
nostrip = {'pysam'}
if name in nostrip:
try:
_shutil.rmtree(archive.getnames()[0])
except OSError:
pass
#_shutil.rmtree(_os.path.sep.join([destination, archive.getnames()[0]]))
# some python modules should not be stripped . . more complex install
for member in archive.getmembers():
if member.isreg():
archive.extract(member)
print(member.name)
c += 1
else:
# others don't need additional compilation
check_path1 = '{}/{}'.format(release, name)
for member in archive.getmembers():
if member.isreg() and check_path1 in member.name:
member.name = _os.path.sep.join(
member.name.split(_os.path.sep)[1:])
archive.extract(member)
c += 1
print('Extracted {} files'.format(c))
else:
# extract as a generic external program
archive.extractall()
if preparation:
for do_this in preparation:
if 'just_packages' in do_this['arguments']:
# this is the only thing that differentiates this prepare()
# from others that need some chdir <== this should be improved
# see dep dict
curdir = _os.path.abspath(_os.curdir)
_os.chdir(_os.path.pardir)
do_this['function'](*do_this['arguments']['package_list'])
# return to previous folder
_os.chdir(curdir)
else:
extracted_base_dir = archive.getnames()[0].split(
_os.path.sep)[0]
curdir = _os.path.abspath(_os.curdir)
# go to installed folder
_os.chdir(_os.path.sep.join([destination, extracted_base_dir]))
do_this['function'](**do_this['arguments'])
# return to previous folder
_os.chdir(curdir)
_os.chdir(initialdir)
def SPAdes(self,
exe = [],
output_folder = ['assemblies','SPAdes'],
mem_num_gigs = 8,
max_cpus = -1,
single_assembly = False,
careful = True,
only_assembler = False):
'''
de novo assembly of short reads using SPAdes
By default, the provided short reads in dictionary: self.paths_to_reads
will be assembled separately, unless single_assembly set to True in
which case each set of paired read fastq files will be used in a
single assembly.
http://spades.bioinf.spbau.ru/release3.6.1/manual.html
relevent inputs:
-o <output_dir> Specify the output directory. Required option.
--sc required for MDA (single-cell) data.
--only-error-correction
--only-assembler
--careful reduce the number of mismatches and short indels. Run MismatchCorrector – a post processing tool. Recommended.
--continue from the specified output folder starting from the last available check-point
--restart-from <check_point>
ec start from error correction
as restart assembly module from the first iteration
k<int> restart from the iteration with specified k values, e.g. k55
mc restart mismatch correction
--pe1-12 <file_name> interlaced forward and reverse paired-end reads.
--pe1-1 <file_name> File with forward reads.
--pe1-2 <file_name> File with reverse reads.
--pe1-s <file_name> File with unpaired reads . . use --pe2-... for next library
--threads <int>
--memory <int> max memory in Gb
-k <int,int,...> Comma-separated list of odd ascending k-mers
If --sc is set the default value are 21,33,55, for multicell data sets it is auto
--cov-cutoff <float> positive float value, or 'auto', or 'off'. Default value is 'off'
'''
assert isinstance(output_folder, list), 'Provide output folder as list of folders forming path'
base_output_path = _os.path.sep.join(output_folder)
if not _os.path.exists(base_output_path):
_os.makedirs(base_output_path)
# max threads is slightly different to cpus
# . . can probably use more
max_processes = _decide_max_processes( max_cpus )
# if an exe is not provided, use that stored in Dependencies
if len(exe):
use_exe = _os.path.sep.join(exe)
else:
from baga import Dependencies
use_exe = _get_exe_path('spades')
def run_SPAdes(cmd):
proc = _subprocess.Popen(cmd, stdout=_subprocess.PIPE, stderr=_subprocess.PIPE)
# allow for failed SPAdes runs (possibly caused by small fastq files) <== but also check they were actually built properly
try:
stdout_value, stderr_value = proc.communicate()
checkthese = []
getline = False
for line in stdout_value.split('\n'):
if 'Warnings saved to' in line:
getline = False
if getline:
l = line.rstrip()
if len(l):
checkthese += [l]
if 'SPAdes pipeline finished WITH WARNINGS!' in line:
getline = True
if len(checkthese):
print('SPAdes completed with warnings:\n{}\n'.format('\n'.join(checkthese)))
else:
print('SPAdes completed without warnings')
# with open('___SPAdes_{}_good_{}.log'.format(cnum, thetime), 'w') as fout:
# fout.write(stdout_value)
path2contigs = _os.path.sep.join([this_output_path,'contigs.fasta'])
except _subprocess.CalledProcessError as e:
print('SPAdes probably did not complete: error returned ({})'.format(proc.returncode))
print('Error: {}'.format(e))
print('Writing some info relevent to SPAdes crash to ___SPAdes_{}_bad_{}.log'.format(cnum, thetime))
with open('___SPAdes_{}_bad_{}.log'.format(cnum, thetime), 'w') as fout:
fout.write(dir(proc))
fout.write('\n' + str(e.returncode) + '\n')
fout.write(_os.path.sep.join([this_output_path,'contigs.fasta']))
path2contigs = None
return(path2contigs)
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
contigs = {}
if single_assembly:
print('Combining reads aligned at multiple regions into single assembly')
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
for cnum, (pairname, files) in enumerate(self.read_files.items()):
# allow use of tuples or dicts by converting dicts to lists
if isinstance(files, dict):
use_files = []
for k,v in sorted(files.items()):
use_files += [v]
else:
use_files = files
cmd += ['--pe{}-1'.format(cnum+1), use_files[0]]
cmd += ['--pe{}-2'.format(cnum+1), use_files[1]]
try:
# use unpaired reads if available
cmd += ['--pe{}-s'.format(cnum+1), use_files[2]]
except IndexError:
pass
try:
# add a second library if provided
if isinstance(self.read_files2[pairname], dict):
# if a dict supplied, make it a list
use_files2 = []
for k,v in sorted(self.read_files2[pairname].items()):
use_files2 += [v]
else:
use_files2 = self.read_files2[pairname]
cmd += ['--pe{}-1'.format(cnum+2), use_files2[0]]
cmd += ['--pe{}-2'.format(cnum+2), use_files2[1]]
try:
cmd += ['--pe{}-s'.format(cnum+2), use_files2[2]]
except IndexError:
pass
except AttributeError:
pass
## this isn't very flexible:
# retain <sample>__<genome> from pairname:
# pairname == <sample>__<genome>_<start>-<end>+<padding>
# and replace with multiregion
folder = '{}__{}_{}'.format(pairname.split('__')[0],
pairname.split('__')[1].split('_')[0],
'multi_region')
this_output_path = _os.path.sep.join(output_folder + [folder])
if not _os.path.exists(this_output_path):
_os.makedirs(this_output_path)
cmd += ['-o', this_output_path]
cmd += ['--threads', str(max_processes)]
cmd += ['--memory', str(mem_num_gigs)]
if only_assembler:
cmd += ['--only-assembler']
if careful:
cmd += ['--careful']
thetime = _time.asctime( _time.localtime(_time.time()) )
print('about to launch SPAdes . . . at {}'.format(thetime))
print(' '.join(cmd))
contigs['multi_region'] = run_SPAdes(cmd)
else:
start_time = _time.time()
# prepare commandline and launch each SPAdes assembly
contigs = {}
for cnum, (pairname, files) in enumerate(sorted(self.read_files.items())):
if isinstance(use_exe, list):
# allow for use of prepended executable with script to run
cmd = list(use_exe)
else:
# or just executable
cmd = [use_exe]
# allow use of tuples or dicts by converting dicts to lists
if isinstance(files, dict):
use_files = []
for k,v in sorted(files.items()):
use_files += [v]
else:
use_files = files
cmd += ['--pe1-1', use_files[0]]
cmd += ['--pe1-2', use_files[1]]
try:
# use unpaired reads if available
cmd += ['--pe1-s', use_files[2]]
except IndexError:
pass
try:
# add a second library if provided
if isinstance(self.read_files2[pairname], dict):
# if a dict supplied, make it a list
use_files2 = []
for k,v in sorted(self.read_files2[pairname].items()):
use_files2 += [v]
else:
use_files2 = self.read_files2[pairname]
cmd += ['--pe2-1', use_files2[0]]
cmd += ['--pe2-2', use_files2[1]]
try:
cmd += ['--pe2-s', use_files2[2]]
except IndexError:
pass
except AttributeError:
pass
this_output_path = _os.path.sep.join(output_folder + [pairname])
if not _os.path.exists(this_output_path):
_os.makedirs(this_output_path)
cmd += ['-o', this_output_path]
cmd += ['--threads', str(max_processes)]
cmd += ['--memory', str(mem_num_gigs)]
if only_assembler:
cmd += ['--only-assembler']
if careful:
cmd += ['--careful']
thetime = _time.asctime( _time.localtime(_time.time()) )
print('about to launch SPAdes . . . at {}'.format(thetime))
print(' '.join(cmd))
contigs[pairname] = run_SPAdes(cmd)
if len(self.read_files) > 1:
# report durations, time left etc
_report_time(start_time, cnum, len(self.read_files))
self.paths_to_contigs = contigs
def align(self, insert_size = False,
path_to_exe = False,
local_alns_path = ['alignments'],
force = False,
max_cpus = -1):
if not path_to_exe:
path_to_exe = _get_exe_path('bwa')
# write genome sequence to a fasta file
try:
_os.makedirs('genome_sequences')
except OSError:
pass
genome_fna = 'genome_sequences/%s.fna' % self.genome_id
_SeqIO.write(_SeqRecord(_Seq(self.genome_sequence.tostring()), id = self.genome_id),
genome_fna,
'fasta')
# make folder for alignments (BAMs)
local_alns_path = _os.path.sep.join(local_alns_path)
if not _os.path.exists(local_alns_path):
_os.makedirs(local_alns_path)
# make a subdir for this genome
local_alns_path_genome = _os.path.sep.join([
local_alns_path,
self.genome_id])
if not _os.path.exists(local_alns_path_genome):
_os.makedirs(local_alns_path_genome)
max_processes = _decide_max_processes( max_cpus )
e1 = 'Could not find "read_files" attribute. Before aligning to genome, reads must be quality score trimmed. Please run trim() method on this Reads instance.'
assert hasattr(self, 'read_files'), e1
e2 = 'Could not find %s. Either run trim() again or ensure file exists'
for pairname, files in self.read_files.items():
assert _os.path.exists(files[1]), e2 % files[1]
assert _os.path.exists(files[2]), e2 % files[2]
have_index_files = [_os.path.exists(genome_fna + '.' + a) for a in ('ann','pac','amb','bwt','sa')]
if not all(have_index_files):
print('Writing BWA index files for %s' % genome_fna)
_subprocess.call([path_to_exe, 'index', genome_fna])
aligned_read_files = {}
for pairname,files in self.read_files.items():
RGinfo = r"@RG\tID:%s\tSM:%s\tPL:ILLUMINA" % (pairname,pairname)
if insert_size:
cmd = [path_to_exe, 'mem', '-t', str(max_processes), '-M', '-a', '-I', insert_size, '-R', RGinfo, genome_fna, files[1], files[2]]
else:
# BWA can estimate on-the-fly
cmd = [path_to_exe, 'mem', '-t', str(max_processes), '-M', '-a', '-R', RGinfo, genome_fna, files[1], files[2]]
out_sam = _os.path.sep.join([local_alns_path_genome, '%s__%s.sam' % (pairname, self.genome_id)])
if not _os.path.exists(out_sam) or force:
print('Called: "%s"' % ' '.join(cmd))
with open(out_sam, "wb") as out:
_subprocess.call(cmd, stdout = out)
else:
print('Found:')
print(out_sam)
print('use "force = True" to overwrite')
print(' '.join(cmd))
aligned_read_files[pairname] = out_sam
self.aligned_read_files = aligned_read_files
def align(self,
insert_size=False,
path_to_exe=False,
local_alns_path=['alignments'],
force=False,
max_cpus=-1):
if not path_to_exe:
path_to_exe = _get_exe_path('bwa')
# write genome sequence to a fasta file
try:
_os.makedirs('genome_sequences')
except OSError:
pass
genome_fna = 'genome_sequences/%s.fna' % self.genome_id
_SeqIO.write(
_SeqRecord(_Seq(self.genome_sequence.tostring()),
id=self.genome_id), genome_fna, 'fasta')
# make folder for alignments (BAMs)
local_alns_path = _os.path.sep.join(local_alns_path)
if not _os.path.exists(local_alns_path):
_os.makedirs(local_alns_path)
# make a subdir for this genome
local_alns_path_genome = _os.path.sep.join(
[local_alns_path, self.genome_id])
if not _os.path.exists(local_alns_path_genome):
_os.makedirs(local_alns_path_genome)
max_processes = _decide_max_processes(max_cpus)
e1 = 'Could not find "read_files" attribute. Before aligning to genome, reads must be quality score trimmed. Please run trim() method on this Reads instance.'
assert hasattr(self, 'read_files'), e1
e2 = 'Could not find %s. Either run trim() again or ensure file exists'
for pairname, files in self.read_files.items():
assert _os.path.exists(files[1]), e2 % files[1]
assert _os.path.exists(files[2]), e2 % files[2]
# always (re)index in case of upstream changes in data
print('Writing BWA index files for %s' % genome_fna)
_subprocess.call([path_to_exe, 'index', genome_fna])
aligned_read_files = {}
for pairname, files in self.read_files.items():
RGinfo = r"@RG\tID:%s\tSM:%s\tPL:ILLUMINA" % (pairname, pairname)
if insert_size:
cmd = [
path_to_exe, 'mem', '-t',
str(max_processes), '-M', '-a', '-I', insert_size, '-R',
RGinfo, genome_fna, files[1], files[2]
]
else:
# BWA can estimate on-the-fly
cmd = [
path_to_exe, 'mem', '-t',
str(max_processes), '-M', '-a', '-R', RGinfo, genome_fna,
files[1], files[2]
]
out_sam = _os.path.sep.join([
local_alns_path_genome,
'%s__%s.sam' % (pairname, self.genome_id)
])
if not _os.path.exists(out_sam) or force:
print('Called: "%s"' % ' '.join(cmd))
with open(out_sam, "wb") as out:
_subprocess.call(cmd, stdout=out)
else:
print('Found:')
print(out_sam)
print('use "force = True" to overwrite')
print(' '.join(cmd))
aligned_read_files[pairname] = out_sam
self.aligned_read_files = aligned_read_files
def generateReads(self, path_to_exe = False,
paths_to_genomes = False,
readcov = 60,
readlen = 100,
fraglen = 350,
sterrfraglen = 20,
model = 4,
max_cpus = -1):
'''
Call GemSIM to generate reads
Need to have written genome sequences to generate from, possibly with
generated SNPs, small indels and large deletions.
'''
#max_cpus etc
if paths_to_genomes:
use_genomes = sorted(paths_to_genomes)
elif hasattr(self, 'written_genomes'):
use_genomes = sorted(self.written_genomes)
else:
raise ValueError('provide either paths_to_genomes or generate some then .writeSequences()')
if not path_to_exe:
path_to_exe = _get_exe_path('gemsim')
comment2 = '''
to generate reads put GemSIM v1.6 into subfolder GemSIM_v1.6 and issue these commands:
GemSIM_v1.6/GemReads.py -r LESB58_for_GemSim_01.fasta -n 1980527 -l d -u 350 -s 20 -m GemSIM_v1.6/models/ill100v4_p.gzip -c -q 33 -p -o GemSimLESB58_01
'''
num_pairs = len(self.genome.sequence) * readcov / (readlen*2)
if model == 4:
path_to_model = _os.path.sep.join(path_to_exe.split(_os.path.sep)[:-1] + ['models','ill100v4_p.gzip'])
elif model == 5:
path_to_model = _os.path.sep.join(path_to_exe.split(_os.path.sep)[:-1] + ['models','ill100v5_p.gzip'])
print('Using error model: {}'.format(path_to_model))
print('Generating {:,} {}bp read pairs for {}x coverage depth of a {}bp genome ({})'.format(
num_pairs, readlen, readcov, len(self.genome.sequence), self.genome.id))
processes = set()
max_processes = _decide_max_processes( max_cpus )
import time
start = time.time()
out_raw = []
for i,genome_in in enumerate(use_genomes):
# could use per genome length . . less consistent than using reference
# genome_len = len(_SeqIO.read(genome_in,'fasta').seq)
# num_pairs = genome_len * readcov / (readlen*2)
outprefix = 'GemSim_{}_{:02d}'.format(self.genome.id, i+1)
cmd = [path_to_exe,
'-r', genome_in,
'-n', num_pairs,
'-l', 'd', '-u', fraglen, '-s', sterrfraglen,
'-m', path_to_model,
'-c',
'-q', 33, '-p',
'-o', outprefix]
out_raw += [outprefix+'_fir.fastq', outprefix+'_sec.fastq']
# this would be better to rename and compress all in one
# maybe as a shell script? Then resuming (--force) would be easier.
if _os.path.exists(outprefix+'_fir.fastq') and \
_os.path.exists(outprefix+'_sec.fastq'):
print('Found output for {}_fir.fastq (and sec), not regenerating, '\
'delete these to start from scratch'.format(outprefix))
else:
cmd = map(str,cmd)
print(' '.join(cmd))
processes.add( _subprocess.Popen(cmd, shell=False) )
if len(processes) >= max_processes:
(pid, exit_status) = _os.wait()
processes.difference_update(
[p for p in processes if p.poll() is not None])
# Check if all the child processes were closed
for p in processes:
if p.poll() is None:
p.wait()
missing = []
for o in out_raw:
if not _os.path.exists(o):
missing += [o]
assert len(missing) == 0, 'Could not find:\n{}'.format('\n'.join(missing))
print('all finished after {} minutes'.format(int(round((time.time() - start)/60.0))))
outdir = _os.path.sep.join(['simulated_reads',self.genome.id])
try:
_os.makedirs(outdir)
except OSError:
pass
for o in out_raw:
new = _os.path.sep.join([outdir, o.replace('fir','R1').replace('sec','R2')])
print('{} ==> {}'.format(o, new))
_os.rename(o, new)
cmd = ['gzip', new]
print(' '.join(cmd))
_subprocess.call(cmd)
def get_download(name, description, source, url, commit, checksum, destination, preparation, checker):
'''
Download and unpack a dependancy
'''
##
initialdir = _os.path.abspath(_os.curdir)
try:
_os.chdir(destination)
except OSError:
_os.makedirs(destination)
_os.chdir(destination)
if checksum:
hasher_algorithm = checksum.split('=')[0]
if hasher_algorithm == 'md5':
hasher = _hashlib.md5()
elif hasher_algorithm == 'sha1':
hasher = _hashlib.sha1()
elif hasher_algorithm == 'sha224':
hasher = _hashlib.sha224()
elif hasher_algorithm == 'sha256':
hasher = _hashlib.sha256()
elif hasher_algorithm == 'sha384':
hasher = _hashlib.sha384()
elif hasher_algorithm == 'sha512':
hasher = _hashlib.sha512()
else:
print("{} checksums not implemented in Python's hashlib!".format(hasher_algorithm))
print('Downloading: %s' % url)
req = _urllib2.urlopen(url)
CHUNK = 16 * 1024 * 16
data = _cStringIO.StringIO()
c = 0
for chunk in iter(lambda: req.read(CHUNK), ''):
c += CHUNK
print("{:,} bytes".format(c))
data.write(chunk)
print('Download complete . . .')
data.seek(0)
if checksum:
buff = data.read(65536)
while len(buff) > 0:
hasher.update(buff)
buff = data.read(65536)
e = '. . . checksum fail!'
assert hasher.hexdigest() == checksum.split('=')[1], e
print('. . . checksum passed!')
data.seek(0)
if url[-6:] == 'tar.gz':
archive = _tarfile.open(mode="r:gz", fileobj = data)
elif url[-7:] == 'tar.bz2':
archive = _tarfile.open(mode="r:bz2", fileobj = data)
elif url[-4:] == '.zip':
archive = _zipfile.ZipFile(data)
if destination == 'local_packages':
# extract as a pypi python package
release = url.split('/')[-1][:-7]
print('Extracting {} to {}'.format(release, _os.path.sep.join([destination,name])))
c = 0
nostrip = {'pysam'}
if name in nostrip:
try:
_shutil.rmtree(archive.getnames()[0])
except OSError:
pass
#_shutil.rmtree(_os.path.sep.join([destination, archive.getnames()[0]]))
# some python modules should not be stripped . . more complex install
for member in archive.getmembers():
if member.isreg():
archive.extract(member)
print(member.name)
c += 1
else:
# others don't need additional compilation
check_path1 = '{}/{}'.format(release,name)
for member in archive.getmembers():
if member.isreg() and check_path1 in member.name:
member.name = _os.path.sep.join(member.name.split(_os.path.sep)[1:])
archive.extract(member)
c += 1
print('Extracted {} files'.format(c))
else:
# extract as a generic external program
archive.extractall()
if preparation:
for do_this in preparation:
if 'just_packages' in do_this['arguments']:
# this is the only thing that differentiates this prepare()
# from others that need some chdir <== this should be improved
# see dep dict
curdir = _os.path.abspath(_os.curdir)
_os.chdir(_os.path.pardir)
do_this['function'](*do_this['arguments']['package_list'])
# return to previous folder
_os.chdir(curdir)
else:
extracted_base_dir = archive.getnames()[0].split(_os.path.sep)[0]
curdir = _os.path.abspath(_os.curdir)
# go to installed folder
_os.chdir(_os.path.sep.join([destination,extracted_base_dir]))
do_this['function'](**do_this['arguments'])
# return to previous folder
_os.chdir(curdir)
_os.chdir(initialdir)
def generateReads(self,
path_to_exe=False,
paths_to_genomes=False,
readcov=60,
readlen=100,
fraglen=350,
sterrfraglen=20,
model=4,
max_cpus=-1):
'''
Call GemSIM to generate reads
Need to have written genome sequences to generate from, possibly with
generated SNPs, small indels and large deletions.
'''
#max_cpus etc
if paths_to_genomes:
use_genomes = sorted(paths_to_genomes)
elif hasattr(self, 'written_genomes'):
use_genomes = sorted(self.written_genomes)
else:
raise ValueError(
'provide either paths_to_genomes or generate some then .writeSequences()'
)
if not path_to_exe:
path_to_exe = _get_exe_path('gemsim')
comment2 = '''
to generate reads put GemSIM v1.6 into subfolder GemSIM_v1.6 and issue these commands:
GemSIM_v1.6/GemReads.py -r LESB58_for_GemSim_01.fasta -n 1980527 -l d -u 350 -s 20 -m GemSIM_v1.6/models/ill100v4_p.gzip -c -q 33 -p -o GemSimLESB58_01
'''
num_pairs = len(self.genome.sequence) * readcov / (readlen * 2)
if model == 4:
path_to_model = _os.path.sep.join(
path_to_exe.split(_os.path.sep)[:-1] +
['models', 'ill100v4_p.gzip'])
elif model == 5:
path_to_model = _os.path.sep.join(
path_to_exe.split(_os.path.sep)[:-1] +
['models', 'ill100v5_p.gzip'])
print('Using error model: {}'.format(path_to_model))
print(
'Generating {:,} {}bp read pairs for {}x coverage depth of a {}bp genome ({})'
.format(num_pairs, readlen, readcov, len(self.genome.sequence),
self.genome.id))
processes = set()
max_processes = _decide_max_processes(max_cpus)
import time
start = time.time()
out_raw = []
for i, genome_in in enumerate(use_genomes):
# could use per genome length . . less consistent than using reference
# genome_len = len(_SeqIO.read(genome_in,'fasta').seq)
# num_pairs = genome_len * readcov / (readlen*2)
outprefix = 'GemSim_{}_{:02d}'.format(self.genome.id, i + 1)
cmd = [
path_to_exe, '-r', genome_in, '-n', num_pairs, '-l', 'd', '-u',
fraglen, '-s', sterrfraglen, '-m', path_to_model, '-c', '-q',
33, '-p', '-o', outprefix
]
out_raw += [outprefix + '_fir.fastq', outprefix + '_sec.fastq']
# this would be better to rename and compress all in one
# maybe as a shell script? Then resuming (--force) would be easier.
if _os.path.exists(outprefix+'_fir.fastq') and \
_os.path.exists(outprefix+'_sec.fastq'):
print('Found output for {}_fir.fastq (and sec), not regenerating, '\
'delete these to start from scratch'.format(outprefix))
else:
cmd = map(str, cmd)
print(' '.join(cmd))
processes.add(_subprocess.Popen(cmd, shell=False))
if len(processes) >= max_processes:
(pid, exit_status) = _os.wait()
processes.difference_update(
[p for p in processes if p.poll() is not None])
# Check if all the child processes were closed
for p in processes:
if p.poll() is None:
p.wait()
missing = []
for o in out_raw:
if not _os.path.exists(o):
missing += [o]
assert len(missing) == 0, 'Could not find:\n{}'.format(
'\n'.join(missing))
print('all finished after {} minutes'.format(
int(round((time.time() - start) / 60.0))))
outdir = _os.path.sep.join(['simulated_reads', self.genome.id])
try:
_os.makedirs(outdir)
except OSError:
pass
for o in out_raw:
new = _os.path.sep.join(
[outdir, o.replace('fir', 'R1').replace('sec', 'R2')])
print('{} ==> {}'.format(o, new))
_os.rename(o, new)
cmd = ['gzip', new]
print(' '.join(cmd))
_subprocess.call(cmd)
