def diginorm(args):
"""
%prog diginorm fastqfile
Run K-mer based normalization. Based on tutorial:
<http://ged.msu.edu/angus/diginorm-2012/tutorial.html>
Assume input is either an interleaved pairs file, or two separate files.
To set up khmer:
$ git clone git://github.com/ged-lab/screed.git
$ git clone git://github.com/ged-lab/khmer.git
$ cd screed
$ python setup.py install
$ cd ../khmer
$ make test
$ export PYTHONPATH=/root/khmer/python
"""
from jcvi.formats.fastq import shuffle, pairinplace, split
p = OptionParser(diginorm.__doc__)
p.set_depth()
p.set_home("khmer")
opts, args = p.parse_args(args)
if len(args) not in (1, 2):
sys.exit(not p.print_help())
if len(args) == 2:
fastq = shuffle(args + ["--tag"])
else:
fastq, = args
kh = opts.khmer_home
depth = opts.depth
sys.path.insert(0, op.join(kh, "python"))
pf = fastq.rsplit(".", 1)[0]
hashfile = pf + ".kh"
keepfile = fastq + ".keep"
norm_cmd = op.join(kh, "scripts/normalize-by-median.py")
filt_cmd = op.join(kh, "scripts/filter-abund.py")
if need_update(fastq, (hashfile, keepfile)):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x 2.5e8 -p".format(depth)
cmd += " --savehash {0} {1}".format(hashfile, fastq)
sh(cmd)
abundfiltfile = keepfile + ".abundfilt"
if need_update((hashfile, keepfile), abundfiltfile):
cmd = filt_cmd
cmd += " {0} {1}".format(hashfile, keepfile)
sh(cmd)
seckeepfile = abundfiltfile + ".keep"
if need_update(abundfiltfile, seckeepfile):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x 1e8".format(depth - 5)
cmd += " {0}".format(abundfiltfile)
sh(cmd)
pairsfile = pairinplace([seckeepfile,
"--base={0}".format(pf + "_norm"), "--rclip=2"])
split([pairsfile])
def diginorm(args):
"""
%prog diginorm fastqfile
Run K-mer based normalization. Based on tutorial:
<http://ged.msu.edu/angus/diginorm-2012/tutorial.html>
Assume input is either an interleaved pairs file, or two separate files.
To set up khmer:
$ git clone git://github.com/ged-lab/screed.git
$ git clone git://github.com/ged-lab/khmer.git
$ cd screed
$ python setup.py install
$ cd ../khmer
$ make test
$ export PYTHONPATH=~/export/khmer
"""
from jcvi.formats.fastq import shuffle, pairinplace, split
from jcvi.apps.base import getfilesize
p = OptionParser(diginorm.__doc__)
p.add_option("--single", default=False, action="store_true",
help="Single end reads")
p.add_option("--tablesize", help="Memory size")
p.add_option("--npass", default="1", choices=("1", "2"),
help="How many passes of normalization")
p.set_depth(depth=50)
p.set_home("khmer", default="/usr/local/bin/")
opts, args = p.parse_args(args)
if len(args) not in (1, 2):
sys.exit(not p.print_help())
if len(args) == 2:
fastq = shuffle(args + ["--tag"])
else:
fastq, = args
kh = opts.khmer_home
depth = opts.depth
PE = not opts.single
sys.path.insert(0, op.join(kh, "python"))
pf = fastq.rsplit(".", 1)[0]
keepfile = fastq + ".keep"
hashfile = pf + ".kh"
mints = 10000000
ts = opts.tablesize or ((getfilesize(fastq) / 16 / mints + 1) * mints)
norm_cmd = op.join(kh, "normalize-by-median.py")
filt_cmd = op.join(kh, "filter-abund.py")
if need_update(fastq, (hashfile, keepfile)):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x {1}".format(depth, ts)
if PE:
cmd += " -p"
cmd += " -s {0} {1}".format(hashfile, fastq)
sh(cmd)
abundfiltfile = keepfile + ".abundfilt"
if need_update((hashfile, keepfile), abundfiltfile):
cmd = filt_cmd
cmd += " {0} {1}".format(hashfile, keepfile)
sh(cmd)
if opts.npass == "1":
seckeepfile = abundfiltfile
else:
seckeepfile = abundfiltfile + ".keep"
if need_update(abundfiltfile, seckeepfile):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x {1}".format(depth - 10, ts / 2)
cmd += " {0}".format(abundfiltfile)
sh(cmd)
if PE:
pairsfile = pairinplace([seckeepfile,
"--base={0}".format(pf + "_norm"), "--rclip=2"])
split([pairsfile])
def diginorm(args):
"""
%prog diginorm fastqfile
Run K-mer based normalization. Based on tutorial:
<http://ged.msu.edu/angus/diginorm-2012/tutorial.html>
Assume input is either an interleaved pairs file, or two separate files.
To set up khmer:
$ git clone git://github.com/ged-lab/screed.git
$ git clone git://github.com/ged-lab/khmer.git
$ cd screed
$ python setup.py install
$ cd ../khmer
$ make test
$ export PYTHONPATH=~/export/khmer
"""
from jcvi.formats.fastq import shuffle, pairinplace, split
from jcvi.apps.base import getfilesize
p = OptionParser(diginorm.__doc__)
p.add_option("--single", default=False, action="store_true",
help="Single end reads")
p.add_option("--tablesize", help="Memory size")
p.add_option("--npass", default="1", choices=("1", "2"),
help="How many passes of normalization")
p.set_depth(depth=50)
p.set_home("khmer", default="/usr/local/bin/")
opts, args = p.parse_args(args)
if len(args) not in (1, 2):
sys.exit(not p.print_help())
if len(args) == 2:
fastq = shuffle(args + ["--tag"])
else:
fastq, = args
kh = opts.khmer_home
depth = opts.depth
PE = not opts.single
sys.path.insert(0, op.join(kh, "python"))
pf = fastq.rsplit(".", 1)[0]
keepfile = fastq + ".keep"
hashfile = pf + ".kh"
mints = 10000000
ts = opts.tablesize or ((getfilesize(fastq) / 16 / mints + 1) * mints)
norm_cmd = op.join(kh, "normalize-by-median.py")
filt_cmd = op.join(kh, "filter-abund.py")
if need_update(fastq, (hashfile, keepfile)):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x {1}".format(depth, ts)
if PE:
cmd += " -p"
cmd += " -s {0} {1}".format(hashfile, fastq)
sh(cmd)
abundfiltfile = keepfile + ".abundfilt"
if need_update((hashfile, keepfile), abundfiltfile):
cmd = filt_cmd
cmd += " {0} {1}".format(hashfile, keepfile)
sh(cmd)
if opts.npass == "1":
seckeepfile = abundfiltfile
else:
seckeepfile = abundfiltfile + ".keep"
if need_update(abundfiltfile, seckeepfile):
cmd = norm_cmd
cmd += " -C {0} -k 20 -N 4 -x {1}".format(depth - 10, ts / 2)
cmd += " {0}".format(abundfiltfile)
sh(cmd)
if PE:
pairsfile = pairinplace([seckeepfile,
"--base={0}".format(pf + "_norm"), "--rclip=2"])
split([pairsfile])
