def merger(args):
"""
%prog merger layout gkpStore contigs.fasta
Merge reads into one contig.
"""
p = OptionParser(merger.__doc__)
opts, args = p.parse_args(args)
if len(args) != 3:
sys.exit(not p.print_help())
layout, gkpstore, contigs = args
fp = open(layout)
pf = "0"
iidfile = pf + ".iids"
for i, row in enumerate(fp):
logging.debug("Read unitig {0}".format(i))
fw = open(iidfile, "w")
layout = row.split("|")
print("\n".join(layout), file=fw)
fw.close()
cmd = "gatekeeper -iid {0}.iids -dumpfasta {0} {1}".format(
pf, gkpstore)
sh(cmd)
fastafile = "{0}.fasta".format(pf)
newfastafile = "{0}.new.fasta".format(pf)
format([
fastafile,
newfastafile,
"--sequential=replace",
"--sequentialoffset=1",
"--nodesc",
])
fasta([newfastafile])
sh("rm -rf {0}".format(pf))
cmd = "runCA {0}.frg -p {0} -d {0} consensus=pbutgcns".format(pf)
cmd += " unitigger=bogart doFragmentCorrection=0 doUnitigSplitting=0"
sh(cmd)
outdir = "{0}/9-terminator".format(pf)
cmd = "cat {0}/{1}.ctg.fasta {0}/{1}.deg.fasta {0}/{1}.singleton.fasta".format(
outdir, pf)
sh(cmd, outfile=contigs, append=True)
def build(args):
"""
%prog build current.fasta Bacteria_Virus.fasta prefix
Build assembly files after a set of clean-ups:
1. Use cdhit (100%) to remove duplicate scaffolds
2. Screen against the bacteria and virus database (remove scaffolds 95% id, 50% cov)
3. Mask matches to UniVec_Core
4. Sort by decreasing scaffold sizes
5. Rename the scaffolds sequentially
6. Build the contigs by splitting scaffolds at gaps
7. Rename the contigs sequentially
"""
from jcvi.apps.cdhit import deduplicate
from jcvi.apps.vecscreen import mask
from jcvi.formats.fasta import sort
p = OptionParser(build.__doc__)
p.add_option(
"--nodedup",
default=False,
action="store_true",
help="Do not deduplicate [default: deduplicate]",
)
opts, args = p.parse_args(args)
if len(args) != 3:
sys.exit(not p.print_help())
fastafile, bacteria, pf = args
dd = deduplicate([fastafile, "--pctid=100"
]) if not opts.nodedup else fastafile
screenfasta = screen([dd, bacteria])
tidyfasta = mask([screenfasta])
sortedfasta = sort([tidyfasta, "--sizes"])
scaffoldfasta = pf + ".assembly.fasta"
format([sortedfasta, scaffoldfasta, "--prefix=scaffold_", "--sequential"])
gapsplitfasta = pf + ".gapSplit.fasta"
cmd = "gapSplit -minGap=10 {0} {1}".format(scaffoldfasta, gapsplitfasta)
sh(cmd)
contigsfasta = pf + ".contigs.fasta"
format([gapsplitfasta, contigsfasta, "--prefix=contig_", "--sequential"])
def merger(args):
"""
%prog merger layout gkpStore contigs.fasta
Merge reads into one contig.
"""
p = OptionParser(merger.__doc__)
opts, args = p.parse_args(args)
if len(args) != 3:
sys.exit(not p.print_help())
layout, gkpstore, contigs = args
fp = open(layout)
pf = "0"
iidfile = pf + ".iids"
for i, row in enumerate(fp):
logging.debug("Read unitig {0}".format(i))
fw = open(iidfile, "w")
layout = row.split("|")
print >> fw, "\n".join(layout)
fw.close()
cmd = "gatekeeper -iid {0}.iids -dumpfasta {0} {1}".format(pf, gkpstore)
sh(cmd)
fastafile = "{0}.fasta".format(pf)
newfastafile = "{0}.new.fasta".format(pf)
format([fastafile, newfastafile, "--sequential=replace", \
"--sequentialoffset=1", "--nodesc"])
fasta([newfastafile])
sh("rm -rf {0}".format(pf))
cmd = "runCA {0}.frg -p {0} -d {0} consensus=pbutgcns".format(pf)
cmd += " unitigger=bogart doFragmentCorrection=0 doUnitigSplitting=0"
sh(cmd)
outdir = "{0}/9-terminator".format(pf)
cmd = "cat {0}/{1}.ctg.fasta {0}/{1}.deg.fasta {0}/{1}.singleton.fasta"\
.format(outdir, pf)
sh(cmd, outfile=contigs, append=True)
def build(args):
"""
%prog build current.fasta Bacteria_Virus.fasta prefix
Build assembly files after a set of clean-ups:
1. Use cdhit (100%) to remove duplicate scaffolds
2. Screen against the bacteria and virus database (remove scaffolds 95% id, 50% cov)
3. Mask matches to UniVec_Core
4. Sort by decreasing scaffold sizes
5. Rename the scaffolds sequentially
6. Build the contigs by splitting scaffolds at gaps
7. Rename the contigs sequentially
"""
from jcvi.apps.cdhit import deduplicate
from jcvi.apps.vecscreen import mask
from jcvi.formats.fasta import sort
p = OptionParser(build.__doc__)
p.add_option("--nodedup", default=False, action="store_true",
help="Do not deduplicate [default: deduplicate]")
opts, args = p.parse_args(args)
if len(args) != 3:
sys.exit(not p.print_help())
fastafile, bacteria, pf = args
dd = deduplicate([fastafile, "--pctid=100"]) \
if not opts.nodedup else fastafile
screenfasta = screen([dd, bacteria])
tidyfasta = mask([screenfasta])
sortedfasta = sort([tidyfasta, "--sizes"])
scaffoldfasta = pf + ".assembly.fasta"
format([sortedfasta, scaffoldfasta, "--prefix=scaffold_", "--sequential"])
gapsplitfasta = pf + ".gapSplit.fasta"
cmd = "gapSplit -minGap=10 {0} {1}".format(scaffoldfasta, gapsplitfasta)
sh(cmd)
contigsfasta = pf + ".contigs.fasta"
format([gapsplitfasta, contigsfasta, "--prefix=contig_", "--sequential"])
def overlap(args):
"""
%prog overlap ctgfasta poolfasta
Fish out the sequences in `poolfasta` that overlap with `ctgfasta`.
Mix and combine using `minimus2`.
"""
p = OptionParser(overlap.__doc__)
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ctgfasta, poolfasta = args
prefix = ctgfasta.split(".")[0]
rid = list(Fasta(ctgfasta).iterkeys())
assert len(rid) == 1, "Use overlapbatch() to improve multi-FASTA file"
rid = rid[0]
splitctgfasta = ctgfasta.rsplit(".", 1)[0] + ".split.fasta"
ctgfasta = run_gapsplit(infile=ctgfasta, outfile=splitctgfasta)
# Run BLAST
blastfile = ctgfasta + ".blast"
run_megablast(infile=ctgfasta, outfile=blastfile, db=poolfasta)
# Extract contigs and merge using minimus2
closuredir = prefix + ".closure"
closure = False
if need_update(blastfile, closuredir):
mkdir(closuredir, overwrite=True)
closure = True
if closure:
idsfile = op.join(closuredir, prefix + ".ids")
cmd = "cut -f2 {0} | sort -u".format(blastfile)
sh(cmd, outfile=idsfile)
idsfastafile = op.join(closuredir, prefix + ".ids.fasta")
cmd = "faSomeRecords {0} {1} {2}".format(poolfasta, idsfile,
idsfastafile)
sh(cmd)
# This step is a hack to weight the bases from original sequences more
# than the pulled sequences, by literally adding another copy to be used
# in consensus calls.
redundantfastafile = op.join(closuredir, prefix + ".redundant.fasta")
format([ctgfasta, redundantfastafile, "--prefix=RED."])
mergedfastafile = op.join(closuredir, prefix + ".merged.fasta")
cmd = "cat {0} {1} {2}".format(ctgfasta, redundantfastafile,
idsfastafile)
sh(cmd, outfile=mergedfastafile)
afgfile = op.join(closuredir, prefix + ".afg")
cmd = "toAmos -s {0} -o {1}".format(mergedfastafile, afgfile)
sh(cmd)
cwd = os.getcwd()
os.chdir(closuredir)
cmd = "minimus2 {0} -D REFCOUNT=0".format(prefix)
cmd += " -D OVERLAP=100 -D MINID=98"
sh(cmd)
os.chdir(cwd)
# Analyze output, make sure that:
# + Get the singletons of the original set back
# + Drop any contig that is comprised entirely of pulled set
originalIDs = set(Fasta(ctgfasta).iterkeys())
minimuscontig = op.join(closuredir, prefix + ".contig")
c = ContigFile(minimuscontig)
excludecontigs = set()
for rec in c.iter_records():
reads = set(x.id for x in rec.reads)
if reads.isdisjoint(originalIDs):
excludecontigs.add(rec.id)
logging.debug("Exclude contigs: {0}".\
format(", ".join(sorted(excludecontigs))))
finalfasta = prefix + ".improved.fasta_"
fw = open(finalfasta, "w")
minimusfasta = op.join(closuredir, prefix + ".fasta")
f = Fasta(minimusfasta)
for id, rec in f.iteritems_ordered():
if id in excludecontigs:
continue
SeqIO.write([rec], fw, "fasta")
singletonfile = op.join(closuredir, prefix + ".singletons")
singletons = set(x.strip() for x in open(singletonfile))
leftovers = singletons & originalIDs
logging.debug("Pull leftover singletons: {0}".\
format(", ".join(sorted(leftovers))))
f = Fasta(ctgfasta)
for id, rec in f.iteritems_ordered():
if id not in leftovers:
continue
SeqIO.write([rec], fw, "fasta")
fw.close()
fastafile = finalfasta
finalfasta = fastafile.rstrip("_")
format([
fastafile, finalfasta, "--sequential", "--pad0=3",
"--prefix={0}_".format(rid)
])
logging.debug("Improved FASTA written to `{0}`.".format(finalfasta))
n50([ctgfasta])
n50([finalfasta])
errlog = "error.log"
for f in (fastafile, blastfile, errlog):
if op.exists(f):
os.remove(f)
def novo(args):
"""
%prog novo reads.fastq
Reference-free tGBS pipeline.
"""
from jcvi.assembly.kmer import jellyfish, histogram
from jcvi.assembly.preprocess import diginorm
from jcvi.formats.fasta import filter as fasta_filter, format
from jcvi.apps.cdhit import filter as cdhit_filter
p = OptionParser(novo.__doc__)
p.add_option("--technology", choices=("illumina", "454", "iontorrent"),
default="iontorrent", help="Sequencing platform")
p.add_option("--dedup", choices=("uclust", "cdhit"),
default="cdhit", help="Dedup algorithm")
p.set_depth(depth=50)
p.set_align(pctid=96)
p.set_home("cdhit", default="/usr/local/bin/")
p.set_home("fiona", default="/usr/local/bin/")
p.set_home("jellyfish", default="/usr/local/bin/")
p.set_cpus()
opts, args = p.parse_args(args)
if len(args) != 1:
sys.exit(not p.print_help())
fastqfile, = args
cpus = opts.cpus
depth = opts.depth
pf, sf = fastqfile.rsplit(".", 1)
diginormfile = pf + ".diginorm." + sf
if need_update(fastqfile, diginormfile):
diginorm([fastqfile, "--single", "--depth={0}".format(depth)])
keepabund = fastqfile + ".keep.abundfilt"
sh("cp -s {0} {1}".format(keepabund, diginormfile))
jf = pf + "-K23.histogram"
if need_update(diginormfile, jf):
jellyfish([diginormfile, "--prefix={0}".format(pf),
"--cpus={0}".format(cpus),
"--jellyfish_home={0}".format(opts.jellyfish_home)])
genomesize = histogram([jf, pf, "23"])
fiona = pf + ".fiona.fa"
if need_update(diginormfile, fiona):
cmd = op.join(opts.fiona_home, "fiona")
cmd += " -g {0} -nt {1} --sequencing-technology {2}".\
format(genomesize, cpus, opts.technology)
cmd += " -vv {0} {1}".format(diginormfile, fiona)
logfile = pf + ".fiona.log"
sh(cmd, outfile=logfile, errfile=logfile)
dedup = opts.dedup
pctid = opts.pctid
cons = fiona + ".P{0}.{1}.consensus.fasta".format(pctid, dedup)
if need_update(fiona, cons):
if dedup == "cdhit":
deduplicate([fiona, "--consensus", "--reads",
"--pctid={0}".format(pctid),
"--cdhit_home={0}".format(opts.cdhit_home)])
else:
uclust([fiona, "--pctid={0}".format(pctid)])
filteredfile = pf + ".filtered.fasta"
if need_update(cons, filteredfile):
covfile = pf + ".cov.fasta"
cdhit_filter([cons, "--outfile={0}".format(covfile),
"--minsize={0}".format(depth / 5)])
fasta_filter([covfile, "50", "--outfile={0}".format(filteredfile)])
finalfile = pf + ".final.fasta"
if need_update(filteredfile, finalfile):
format([filteredfile, finalfile, "--sequential=replace",
"--prefix={0}_".format(pf)])
def patch(args):
"""
%prog patch reference.fasta reads.fasta
Run PBJelly with reference and reads.
"""
from jcvi.formats.base import write_file
from jcvi.formats.fasta import format
p = OptionParser(patch.__doc__)
p.add_option("--cleanfasta", default=False, action="store_true",
help="Clean FASTA to remove description [default: %default]")
p.add_option("--highqual", default=False, action="store_true",
help="Reads are of high quality [default: %default]")
p.set_home("pbjelly")
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ref, reads = args
cmd = op.join(opts.pbjelly_home, "setup.sh")
if not which("fakeQuals.py"):
setup = "source {0}".format(cmd)
sh(setup)
# Check environment
try:
import networkx
version = networkx.version
except:
logging.error("You need networkx==1.1 to run PBJELLY")
return
try:
import argparse
except ImportError:
logging.error("You need Python2.7 or at least argparse lib")
return
pf = ref.rsplit(".", 1)[0]
pr, px = reads.rsplit(".", 1)
# Remove description line
if opts.cleanfasta:
oref = pf + ".f.fasta"
oreads = pr + ".f.fasta"
format([ref, oref])
format([reads, oreads])
ref, reads = oref, oreads
# Check if the FASTA has qual
ref, refq = fake_quals(ref)
convert_reads = not px in ("fq", "fastq", "txt")
if convert_reads:
reads, readsq = fake_quals(reads)
readsfiles = " ".join((reads, readsq))
else:
readsfiles = reads
# Make directory structure
dref, dreads = "data/reference", "data/reads"
sh("mkdir -p {0}".format(dref))
sh("mkdir -p {0}".format(dreads))
sh("cp {0} {1}/".format(" ".join((ref, refq)), dref))
sh("cp {0} {1}/".format(readsfiles, dreads))
cwd = os.getcwd()
outputDir = cwd
reference = op.join(cwd, "{0}/{1}".format(dref, ref))
reads = op.join(cwd, "{0}/{1}".format(dreads, reads))
p = Protocol(outputDir, reference, reads, highqual=opts.highqual)
p.write_xml()
# Make sure we have the patched version of Extraction.py
# See discussion <http://seqanswers.com/forums/showthread.php?t=27599>
# This check has been removed
# Build the pipeline
runsh = [setup]
for action in "setup|mapping|support|extraction".split("|"):
runsh.append("Jelly.py {0} Protocol.xml".format(action))
#pcmds = """find assembly -name "ref*" -exec echo \\
# "Assembly.py {} \\
# > {}/assembly.out 2> {}/assembly.err" \; > commands.list"""
#runsh.append(pcmds)
runsh.append("Jelly.py assembly Protocol.xml")
runsh.append("cp assembly/assembly_chunk0.sh commands.list")
runsh.append("parallel < commands.list")
runsh.append("Jelly.py output Protocol.xml")
runfile = "run.sh"
contents = "\n".join(runsh)
write_file(runfile, contents)
def patch(args):
"""
%prog patch reference.fasta reads.fasta
Run PBJelly with reference and reads.
"""
from jcvi.formats.base import write_file
from jcvi.formats.fasta import format
p = OptionParser(patch.__doc__)
p.add_option("--cleanfasta", default=False, action="store_true",
help="Clean FASTA to remove description [default: %default]")
p.add_option("--highqual", default=False, action="store_true",
help="Reads are of high quality [default: %default]")
p.set_home("pbjelly")
p.set_cpus()
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ref, reads = args
cpus = opts.cpus
cmd = op.join(opts.pbjelly_home, "setup.sh")
setup = "source {0}".format(cmd)
if not which("fakeQuals.py"):
sh(setup)
pf = ref.rsplit(".", 1)[0]
pr, px = reads.rsplit(".", 1)
# Remove description line
if opts.cleanfasta:
oref = pf + ".f.fasta"
oreads = pr + ".f.fasta"
format([ref, oref])
format([reads, oreads])
ref, reads = oref, oreads
# Check if the FASTA has qual
ref, refq = fake_quals(ref)
convert_reads = not px in ("fq", "fastq", "txt")
if convert_reads:
reads, readsq = fake_quals(reads)
readsfiles = " ".join((reads, readsq))
else:
readsfiles = reads
# Make directory structure
dref, dreads = "data/reference", "data/reads"
cwd = os.getcwd()
reference = op.join(cwd, "{0}/{1}".format(dref, ref))
reads = op.join(cwd, "{0}/{1}".format(dreads, reads))
if not op.exists(reference):
sh("mkdir -p {0}".format(dref))
sh("cp {0} {1}/".format(" ".join((ref, refq)), dref))
if not op.exists(reads):
sh("mkdir -p {0}".format(dreads))
sh("cp {0} {1}/".format(readsfiles, dreads))
outputDir = cwd
p = Protocol(outputDir, reference, reads, highqual=opts.highqual)
p.write_xml()
# Build the pipeline
runsh = [setup]
for action in "setup|mapping|support|extraction".split("|"):
runsh.append("Jelly.py {0} Protocol.xml".format(action))
runsh.append('Jelly.py assembly Protocol.xml -x "--nproc={0}"'.format(cpus))
runsh.append("Jelly.py output Protocol.xml")
runfile = "run.sh"
contents = "\n".join(runsh)
write_file(runfile, contents)
def patch(args):
"""
%prog patch reference.fasta reads.fasta
Run PBJelly with reference and reads.
"""
from jcvi.formats.base import write_file
from jcvi.formats.fasta import format
p = OptionParser(patch.__doc__)
p.add_option("--cleanfasta",
default=False,
action="store_true",
help="Clean FASTA to remove description [default: %default]")
p.add_option("--highqual",
default=False,
action="store_true",
help="Reads are of high quality [default: %default]")
p.set_home("pbjelly")
p.set_cpus()
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ref, reads = args
cpus = opts.cpus
cmd = op.join(opts.pbjelly_home, "setup.sh")
setup = "source {0}".format(cmd)
if not which("fakeQuals.py"):
sh(setup)
pf = ref.rsplit(".", 1)[0]
pr, px = reads.rsplit(".", 1)
# Remove description line
if opts.cleanfasta:
oref = pf + ".f.fasta"
oreads = pr + ".f.fasta"
format([ref, oref])
format([reads, oreads])
ref, reads = oref, oreads
# Check if the FASTA has qual
ref, refq = fake_quals(ref)
convert_reads = not px in ("fq", "fastq", "txt")
if convert_reads:
reads, readsq = fake_quals(reads)
readsfiles = " ".join((reads, readsq))
else:
readsfiles = reads
# Make directory structure
dref, dreads = "data/reference", "data/reads"
cwd = os.getcwd()
reference = op.join(cwd, "{0}/{1}".format(dref, ref))
reads = op.join(cwd, "{0}/{1}".format(dreads, reads))
if not op.exists(reference):
sh("mkdir -p {0}".format(dref))
sh("cp {0} {1}/".format(" ".join((ref, refq)), dref))
if not op.exists(reads):
sh("mkdir -p {0}".format(dreads))
sh("cp {0} {1}/".format(readsfiles, dreads))
outputDir = cwd
p = Protocol(outputDir, reference, reads, highqual=opts.highqual)
p.write_xml()
# Build the pipeline
runsh = [setup]
for action in "setup|mapping|support|extraction".split("|"):
runsh.append("Jelly.py {0} Protocol.xml".format(action))
runsh.append(
'Jelly.py assembly Protocol.xml -x "--nproc={0}"'.format(cpus))
runsh.append("Jelly.py output Protocol.xml")
runfile = "run.sh"
contents = "\n".join(runsh)
write_file(runfile, contents)
def overlap(args):
"""
%prog overlap ctgfasta poolfasta
Fish out the sequences in `poolfasta` that overlap with `ctgfasta`.
Mix and combine using `minimus2`.
"""
p = OptionParser(overlap.__doc__)
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ctgfasta, poolfasta = args
prefix = ctgfasta.split(".")[0]
rid = list(Fasta(ctgfasta).iterkeys())
assert len(rid) == 1, "Use overlapbatch() to improve multi-FASTA file"
rid = rid[0]
splitctgfasta = ctgfasta.rsplit(".", 1)[0] + ".split.fasta"
ctgfasta = run_gapsplit(infile=ctgfasta, outfile=splitctgfasta)
# Run BLAST
blastfile = ctgfasta + ".blast"
run_megablast(infile=ctgfasta, outfile=blastfile, db=poolfasta)
# Extract contigs and merge using minimus2
closuredir = prefix + ".closure"
closure = False
if need_update(blastfile, closuredir):
mkdir(closuredir, overwrite=True)
closure = True
if closure:
idsfile = op.join(closuredir, prefix + ".ids")
cmd = "cut -f2 {0} | sort -u".format(blastfile)
sh(cmd, outfile=idsfile)
idsfastafile = op.join(closuredir, prefix + ".ids.fasta")
cmd = "faSomeRecords {0} {1} {2}".format(poolfasta, idsfile, idsfastafile)
sh(cmd)
# This step is a hack to weight the bases from original sequences more
# than the pulled sequences, by literally adding another copy to be used
# in consensus calls.
redundantfastafile = op.join(closuredir, prefix + ".redundant.fasta")
format([ctgfasta, redundantfastafile, "--prefix=RED."])
mergedfastafile = op.join(closuredir, prefix + ".merged.fasta")
cmd = "cat {0} {1} {2}".format(ctgfasta, redundantfastafile, idsfastafile)
sh(cmd, outfile=mergedfastafile)
afgfile = op.join(closuredir, prefix + ".afg")
cmd = "toAmos -s {0} -o {1}".format(mergedfastafile, afgfile)
sh(cmd)
cwd = os.getcwd()
os.chdir(closuredir)
cmd = "minimus2 {0} -D REFCOUNT=0".format(prefix)
cmd += " -D OVERLAP=100 -D MINID=98"
sh(cmd)
os.chdir(cwd)
# Analyze output, make sure that:
# + Get the singletons of the original set back
# + Drop any contig that is comprised entirely of pulled set
originalIDs = set(Fasta(ctgfasta).iterkeys())
minimuscontig = op.join(closuredir, prefix + ".contig")
c = ContigFile(minimuscontig)
excludecontigs = set()
for rec in c.iter_records():
reads = set(x.id for x in rec.reads)
if reads.isdisjoint(originalIDs):
excludecontigs.add(rec.id)
logging.debug("Exclude contigs: {0}".\
format(", ".join(sorted(excludecontigs))))
finalfasta = prefix + ".improved.fasta_"
fw = open(finalfasta, "w")
minimusfasta = op.join(closuredir, prefix + ".fasta")
f = Fasta(minimusfasta)
for id, rec in f.iteritems_ordered():
if id in excludecontigs:
continue
SeqIO.write([rec], fw, "fasta")
singletonfile = op.join(closuredir, prefix + ".singletons")
singletons = set(x.strip() for x in open(singletonfile))
leftovers = singletons & originalIDs
logging.debug("Pull leftover singletons: {0}".\
format(", ".join(sorted(leftovers))))
f = Fasta(ctgfasta)
for id, rec in f.iteritems_ordered():
if id not in leftovers:
continue
SeqIO.write([rec], fw, "fasta")
fw.close()
fastafile = finalfasta
finalfasta = fastafile.rstrip("_")
format([fastafile, finalfasta, "--sequential", "--pad0=3",
"--prefix={0}_".format(rid)])
logging.debug("Improved FASTA written to `{0}`.".format(finalfasta))
n50([ctgfasta])
n50([finalfasta])
errlog = "error.log"
for f in (fastafile, blastfile, errlog):
if op.exists(f):
os.remove(f)
def novo(args):
"""
%prog novo reads.fastq
Reference-free tGBS pipeline.
"""
from jcvi.assembly.kmer import jellyfish, histogram
from jcvi.assembly.preprocess import diginorm
from jcvi.formats.fasta import filter as fasta_filter, format
from jcvi.apps.cdhit import filter as cdhit_filter
p = OptionParser(novo.__doc__)
p.add_option("--technology",
choices=("illumina", "454", "iontorrent"),
default="iontorrent",
help="Sequencing platform")
p.add_option("--dedup",
choices=("uclust", "cdhit"),
default="cdhit",
help="Dedup algorithm")
p.set_depth(depth=50)
p.set_align(pctid=96)
p.set_home("cdhit")
p.set_home("fiona")
p.set_cpus()
opts, args = p.parse_args(args)
if len(args) != 1:
sys.exit(not p.print_help())
fastqfile, = args
cpus = opts.cpus
depth = opts.depth
pf, sf = fastqfile.rsplit(".", 1)
diginormfile = pf + ".diginorm." + sf
if need_update(fastqfile, diginormfile):
diginorm([fastqfile, "--single", "--depth={0}".format(depth)])
keepabund = fastqfile + ".keep.abundfilt"
sh("cp -s {0} {1}".format(keepabund, diginormfile))
jf = pf + "-K23.histogram"
if need_update(diginormfile, jf):
jellyfish([
diginormfile, "--prefix={0}".format(pf), "--cpus={0}".format(cpus)
])
genomesize = histogram([jf, pf, "23"])
fiona = pf + ".fiona.fa"
if need_update(diginormfile, fiona):
cmd = op.join(opts.fiona_home, "bin/fiona")
cmd += " -g {0} -nt {1} --sequencing-technology {2}".\
format(genomesize, cpus, opts.technology)
cmd += " -vv {0} {1}".format(diginormfile, fiona)
logfile = pf + ".fiona.log"
sh(cmd, outfile=logfile, errfile=logfile)
dedup = opts.dedup
pctid = opts.pctid
cons = fiona + ".P{0}.{1}.consensus.fasta".format(pctid, dedup)
if need_update(fiona, cons):
if dedup == "cdhit":
deduplicate([
fiona, "--consensus", "--reads", "--pctid={0}".format(pctid),
"--cdhit_home={0}".format(opts.cdhit_home)
])
else:
uclust([fiona, "--pctid={0}".format(pctid)])
filteredfile = pf + ".filtered.fasta"
if need_update(cons, filteredfile):
covfile = pf + ".cov.fasta"
cdhit_filter([
cons, "--outfile={0}".format(covfile),
"--minsize={0}".format(depth / 5)
])
fasta_filter([covfile, "50", "--outfile={0}".format(filteredfile)])
finalfile = pf + ".final.fasta"
if need_update(filteredfile, finalfile):
format([
filteredfile, finalfile, "--sequential=replace",
"--prefix={0}_".format(pf)
])
def patch(args):
"""
%prog patch reference.fasta reads.fasta
Run PBJelly with reference and reads.
"""
from jcvi.formats.base import write_file
from jcvi.formats.fasta import format
p = OptionParser(patch.__doc__)
p.add_option("--cleanfasta", default=False, action="store_true",
help="Clean FASTA to remove description [default: %default]")
p.add_option("--highqual", default=False, action="store_true",
help="Reads are of high quality [default: %default]")
p.set_home("pbjelly")
opts, args = p.parse_args(args)
if len(args) != 2:
sys.exit(not p.print_help())
ref, reads = args
cmd = op.join(opts.pbjelly_home, "setup.sh")
if not which("fakeQuals.py"):
setup = "source {0}".format(cmd)
sh(setup)
# Check environment
try:
import networkx
version = networkx.version
except:
logging.error("You need networkx==1.1 to run PBJELLY")
return
try:
import argparse
except ImportError:
logging.error("You need Python2.7 or at least argparse lib")
return
pf = ref.rsplit(".", 1)[0]
pr, px = reads.rsplit(".", 1)
# Remove description line
if opts.cleanfasta:
oref = pf + ".f.fasta"
oreads = pr + ".f.fasta"
format([ref, oref])
format([reads, oreads])
ref, reads = oref, oreads
# Check if the FASTA has qual
ref, refq = fake_quals(ref)
convert_reads = not px in ("fq", "fastq", "txt")
if convert_reads:
reads, readsq = fake_quals(reads)
readsfiles = " ".join((reads, readsq))
else:
readsfiles = reads
# Make directory structure
dref, dreads = "data/reference", "data/reads"
sh("mkdir -p {0}".format(dref))
sh("mkdir -p {0}".format(dreads))
sh("cp {0} {1}/".format(" ".join((ref, refq)), dref))
sh("cp {0} {1}/".format(readsfiles, dreads))
cwd = os.getcwd()
outputDir = cwd
reference = op.join(cwd, "{0}/{1}".format(dref, ref))
reads = op.join(cwd, "{0}/{1}".format(dreads, reads))
p = Protocol(outputDir, reference, reads, highqual=opts.highqual)
p.write_xml()
# Make sure we have the patched version of Extraction.py
# See discussion <http://seqanswers.com/forums/showthread.php?t=27599>
# This check has been removed
# Build the pipeline
runsh = [setup]
for action in "setup|mapping|support|extraction".split("|"):
runsh.append("Jelly.py {0} Protocol.xml".format(action))
#pcmds = """find assembly -name "ref*" -exec echo \\
# "Assembly.py {} \\
# > {}/assembly.out 2> {}/assembly.err" \; > commands.list"""
#runsh.append(pcmds)
runsh.append("Jelly.py assembly Protocol.xml")
runsh.append("cp assembly/assembly_chunk0.sh commands.list")
runsh.append("parallel < commands.list")
runsh.append("Jelly.py output Protocol.xml")
runfile = "run.sh"
contents = "\n".join(runsh)
write_file(runfile, contents, meta="run script")
