def skimReadGroup(self):
handlers = []
for fn in self.fileNames:
handlers.append(AlignmentFileStack(fn, self.maxEntries))
alignCon = AlignmentConnector()
ovlcount = 0
readCount = 0
while True:
availNames = []
delete = []
for i in handlers:
success = i.gleam() #grabs the next read's set of hits
if success is not None:
availNames.append(success)
else:
delete.append(i)
#get rid of the finished ones
for d in delete:
logging.info("Removing %d fileHandlers" % (len(delete)))
handlers.remove(d)
if len(availNames) == 0:
logging.info("No Reads Left")
return
leastName = natural_sort(availNames)[0]
readGroup = []
for i in handlers:
if i.stack[0].qname == leastName:
if i.stack[0].qseqlength < self.lengthMin:
i.stack = []
readCount += 1
continue
for read in i.stack:
#filtering
#The not [option] or [test] structure
#says if this option is not on, short circuit and don't goto or.
if (read.tseqlength >= self.lengthMin) and \
(not self.extends or alignCon.extendsTarget(read) not in \
[SUPPORTFLAGS.none, SUPPORTFLAGS.span]) and \
(not self.tailMax >= 0 or not \
alignCon.isDiscordant(read, self.tailMax)):
readGroup.append(read)
#reset handler's stack
i.stack = []
readCount += 1
if len(readGroup) > 0:
self.makeTableEntry(readGroup)
if readCount % 10000 == 0:
logging.info("%d reads parsed" % (readCount))
if len(handlers) == 0:
logging.info("Out of File Handlers")
return
def skimReadGroup(self):
handlers = []
for fn in self.fileNames:
handlers.append(AlignmentFileStack(fn, self.maxEntries))
alignCon = AlignmentConnector()
ovlcount = 0
readCount = 0
while True:
availNames = []
delete = []
for i in handlers:
success = i.gleam()#grabs the next read's set of hits
if success is not None:
availNames.append(success)
else:
delete.append(i)
#get rid of the finished ones
for d in delete:
logging.info("Removing %d fileHandlers" % (len(delete)))
handlers.remove(d)
if len(availNames) == 0:
logging.info("No Reads Left")
return
leastName = natural_sort(availNames)[0]
readGroup = []
for i in handlers:
if i.stack[0].qname == leastName:
if i.stack[0].qseqlength < self.lengthMin:
i.stack = []
readCount += 1
continue
for read in i.stack:
#filtering
#The not [option] or [test] structure
#says if this option is not on, short circuit and don't goto or.
if (read.tseqlength >= self.lengthMin) and \
(not self.extends or alignCon.extendsTarget(read) not in \
[SUPPORTFLAGS.none, SUPPORTFLAGS.span]) and \
(not self.tailMax >= 0 or not \
alignCon.isDiscordant(read, self.tailMax)):
readGroup.append(read)
#reset handler's stack
i.stack = []
readCount += 1
if len(readGroup) > 0:
self.makeTableEntry(readGroup)
if readCount % 10000 == 0:
logging.info("%d reads parsed" % (readCount))
if len(handlers) == 0:
logging.info("Out of File Handlers")
return
def singleOverlapAssembly(alldata, args):
"""
"""
global ALLTEMPFILES
data = alldata.stats
reads = NamedTemporaryFile(prefix="sol_", suffix=".fasta", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(reads.name)
e1Seq = data["extendSeq1"]; e2Seq = data["extendSeq2"]
reads.write(">%s\n%s\n>%s\n%s\n" % ("seq1", e1Seq, "seq2", e2Seq))
reads.close()
alignFn = NamedTemporaryFile(prefix="sol_",suffix=".m5", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(alignFn.name)
blasr(reads.name, reads.name, nproc=args.nproc, outname=alignFn.name)
aligns = M5File(alignFn)
# find best hit between the two
connector = AlignmentConnector()
bestS = None
bestA = 0
for i in aligns:
if i.qname != i.tname:
if connector.extendsTarget(i):
if i.score < bestS:
bestA = i
bestS = i.score
if bestS is None:
logging.info("no overlap between extenders")
return
#any of these steps could fail --
#Ensure the hit is valid
#(if + + and sameStrand we are okay, if - + and not sameStrand we are okay)
if data["sameStrand"] == (bestA.tstrand == '0'):
logging.info("bad overlap between extenders")
return
con = consensus([bestA])
bestA = bestA[0]
#strand correction...
if bestA.qname == "seq1":
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.tstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.qstart] + con.sequence.translate(revComp)[::-1] + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tend:]
else:
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.qstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.tstart] + con.sequence + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tstart:]
return seq
def singleOverlapAssembly(alldata, args):
"""
"""
global ALLTEMPFILES
data = alldata.stats
reads = NamedTemporaryFile(prefix="sol_", suffix=".fasta", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(reads.name)
e1Seq = data["extendSeq1"]; e2Seq = data["extendSeq2"]
reads.write(">%s\n%s\n>%s\n%s\n" % ("seq1", e1Seq, "seq2", e2Seq))
reads.close()
alignFn = NamedTemporaryFile(prefix="sol_",suffix=".m5", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(alignFn.name)
blasr(reads.name, reads.name, nproc=args.nproc, outname=alignFn.name)
aligns = M5File(alignFn)
# find best hit between the two
connector = AlignmentConnector()
bestS = None
bestA = 0
for i in aligns:
if i.qname != i.tname:
if connector.extendsTarget(i):
if i.score < bestS:
bestA = i
bestS = i.score
if bestS is None:
logging.info("no overlap between extenders")
return
#any of these steps could fail --
#Ensure the hit is valid
#(if + + and sameStrand we are okay, if - + and not sameStrand we are okay)
if data["sameStrand"] == (bestA.tstrand == '0'):
logging.info("bad overlap between extenders")
return
con = consensus([bestA])
bestA = bestA[0]
#strand correction...
if bestA.qname == "seq1":
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.tstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.qstart] + con.sequence.translate(revComp)[::-1] + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tend:]
else:
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.qstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.tstart] + con.sequence + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tstart:]
return seq
def preunitereads(inputFastq, args):
"""
sent query, I'm going to pop all of the united reads onto this
"""
global ALLTEMPFILES
alignFile = NamedTemporaryFile(prefix="uni_",
suffix=".m5",
delete=False,
dir=args.tempDir).name
ALLTEMPFILES.append(alignFile)
readFile = NamedTemporaryFile(prefix="uni_",
suffix=".fasta",
delete=False,
dir=args.tempDir)
ALLTEMPFILES.append(readFile.name)
input = FastqFile(inputFastq)
for read in input:
readFile.write(">%s\n%s\n" % (input[read].name, input[read].seq))
readFile.close()
readFile = readFile.name
blasr(readFile,
readFile,
bestn=5,
nCandidates=20,
nproc=args.nproc,
outname=alignFile)
aligns = M5File(alignFile)
con = AlignmentConnector()
extenders = []
for a in aligns:
if a.tname == a.qname:
continue
if a.qstart - a.qend < 500 or a.tstart - a.tend < 500:
continue
sup = con.extendsTarget(a, minCovers=500)
#sup = con.extendsTarget(a, minCovers=100)
a.support = sup
if sup in [SUPPORTFLAGS.left, SUPPORTFLAGS.right]:
extenders.append(a)
best = {} #best of queries
for i in extenders:
score = 0
if i.qname in best:
score = best[i.qname].score
if i.score < score:
best[i.qname] = i
#print "q"
#for i in best.values():
#print str(i)
best2 = {} #best of targets
for i in best.values():
score = 0
if i.tname in best2:
score = best2[i.tname].score
if i.score < score:
best2[i.tname] = i
#print "t"
#for i in best2.values():
#print str(i)
best3 = {} #best of both
for i in best2.values():
keys = [i.qname, i.tname]
keys.sort()
keys = "".join(keys)
score = 0
if keys in best3:
score = best3[keys].score
if i.score < score:
best3[keys] = i
#print 'b'
#for i in best3.values():
#print str(i)
reads = FastqFile(inputFastq)
fout = open(inputFastq, 'a')
count = 0
for i in best3.values():
qseq = None
if i.support == SUPPORTFLAGS.left:
if i.qstrand == '0':
qseq = reads[i.qname].seq + reads[i.tname].seq[i.tend:]
elif i.qstrand == '1':
qseq = reads[i.qname].seq + reads[
i.tname].seq[i.tend:].translate(revComp)
if i.support == SUPPORTFLAGS.right:
if i.qstrand == '0':
qseq = reads[i.tname].seq[:i.tstart] + reads[i.qname].seq
elif i.qstrand == '1':
qseq = reads[i.tname].seq[:i.tstart].translate(
revComp) + reads[i.qname].seq
if qseq is not None:
count += 1
fout.write("@%s_%s\n%s\n+\n%s\n" %
(i.qname, i.tname, qseq, "!" * len(qseq)))
logging.info("Preunited %d reads" % (count))
fout.close()
def getSubSeqs(alignmentFile,
readsFile,
sameStrand,
seeds,
predictedGapSize,
maxTrim,
maxWiggle,
basedir="./"):
"""
Finds the seqs that align to the flanks the best, creates a fastq of supporting reads
and the seed
Might have a problem with my best read no going off the edge fully
so I put the maxFlank at 20
I should do more strand correction here
"""
global ALLTEMPFILES
def singleExtendLookup(sup, a):
"""
For getting how a single read extends a single flank
"""
if sup == SUPPORTFLAGS.none:
return None
#set the coordinates of the extending sequence
logging.debug(sup)
logging.debug(a.qname)
mystart = None
myend = None
if a.tname.endswith("e5") and sup in [
SUPPORTFLAGS.left, SUPPORTFLAGS.span
]:
if a.tstrand == '0':
mystart = 0
myend = a.qstart
else:
mystart = a.qend
myend = a.qseqlength
elif a.tname.endswith("e3") and sup in [
SUPPORTFLAGS.right, SUPPORTFLAGS.span
]:
if a.tstrand == '0':
mystart = a.qend
myend = a.qseqlength
else:
mystart = 0
myend = a.qstart
if mystart is None or myend is None or mystart < 0 or myend > a.qseqlength:
return None
#tscore = a.score * (myend - mystart)
#what flank and is it the best
if a.tname.replace('/', '.') == stats["seed1"]:
stats["extendF1Count"] += 1
stats["avgExtF1Bases"] += a.qstart
stats["support"][1].append(sup)
if a.score < stats["extendF1SeedScore"]:
stats["extendF1SeedScore"] = a.score #tscore
stats["extendF1SeedName"] = a.qname
stats["extendF1SeedStart"] = mystart
stats["extendF1SeedEnd"] = myend
stats["extendF1SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f1fout
elif a.tname.replace('/', '.') == stats["seed2"]:
stats["extendF2Count"] += 1
stats["avgExtF2Bases"] += a.qstart
stats["support"][2].append(sup)
if a.score < stats["extendF2SeedScore"]:
stats["extendF2SeedScore"] = a.score #tscore
stats["extendF2SeedName"] = a.qname
stats["extendF2SeedStart"] = mystart
stats["extendF2SeedEnd"] = myend
stats["extendF2SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f2fout
#myOut.write(str(reads[a.qname].subSeq(mystart, myend)))
return None
connector = AlignmentConnector()
#aligns = connector.parseAlignments(M5File(alignmentFile))
#no need to connect with the tailmap
aligns = defaultdict(list)
for a in M4File(alignmentFile):
aligns[a.qname].append(a)
aligns = aligns.values()
reads = FastqFile(readsFile)
stats = createStats()
stats["seed1"], stats["seed2"] = seeds
stats["sameStrand"] = sameStrand
bestSpan = None
bestF1E = None
bestF2E = None
for readGroup in aligns:
if len(readGroup) > 2:
best = 0
worst = 0
keep = []
for i in readGroup:
if i.score < best:
keep.insert(0, i)
if len(keep) >= 2:
keep.pop()
best = i.score
elif i.score < worst:
keep.insert(1, i)
if len(keep) >= 2:
keep.pop()
worst = i.score
readGroup = keep
if len(readGroup) == 2:
#make sure that the two hits aren't hitting the same target
if readGroup[0].tname == readGroup[1].tname:
if readGroup[0].score <= readGroup[1].score:
del (readGroup[1])
else:
del (readGroup[0])
#hit on each flank
if len(readGroup) == 2:
r1, r2 = readGroup
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a = connector.extendsTarget(r1, maxFlank=maxTrim, minCovers=0)
logging.debug(a)
#Also check appropriate orientation
if r1.tname.endswith('e3'):
if a not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
elif r1.tname.endswith('e5'):
if a not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
b = connector.extendsTarget(r2, maxFlank=maxTrim, minCovers=0)
if r2.tname.endswith('e3'):
if b not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif r2.tname.endswith('e5'):
if b not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif len(readGroup) == 1:
r1 = readGroup[0]
r2 = None
a = connector.extendsTarget(r1, maxFlank=10)
b = SUPPORTFLAGS.none
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a, b = b, a
else:
logging.warning("read %s gave too many alignments" %
(readGroup[0].qname))
#it extends both flanks
if a != SUPPORTFLAGS.none and b != SUPPORTFLAGS.none:
logging.debug("%s spans" % r1.qname)
logging.debug("aflag %d bflag %d" % (a, b))
logging.debug("hit1- %s (%d, %d)" % (r1.tname, r1.qstart, r1.qend))
logging.debug("hit2- %s (%d, %d)" % (r2.tname, r2.qstart, r2.qend))
rStart = min(r1.qend, r2.qend)
rEnd = max(r1.qstart, r2.qstart)
sz = rEnd - rStart
tooShort = False
if sz < 50:
logging.info("fill seq is too short to call consensus")
tooShort = True
tooShortSeq = reads[r1.qname].subSeq(rStart, rEnd)
#continue
if predictedGapSize is not None and (predictedGapSize -
sz) > maxWiggle:
logging.info(
"fill seq size %d is smaller than allowed predicted gap size wiggle %d"
% (sz, maxWiggle))
continue
#Need to ensure that it's extending in the correct orientation
#need to ensure that everything is on the right strand
if sameStrand and r1.tstrand != r2.tstrand:
logging.debug("bad strandedness")
continue
#check for negative gaps
stats["spanCount"] += 1
stats["avgSpanBases"] += rEnd - rStart
stats["support"][0].append(SUPPORTFLAGS.span)
t = reads[r1.qname].subSeq(rStart, rEnd)
#sfout.write(str(t))
#is it the best spanner
score = r1.score + r2.score
if score < stats["spanSeedScore"]:
logging.debug("scoring %s %s" % (r1.qname, r2.qname))
stats["spanSeedScore"] = score
spanSeedName = r1.qname
stats["spanSeedStrand1"] = r1.tstrand
bestSpan = reads[r1.qname].subSeq(rStart, rEnd)
stats["spanSeedName"] = r1.qname
stats["spanSeedStart"] = rStart
stats["spanSeedEnd"] = rEnd
stats["spanSeedStrand2"] = r2.tstrand
stats["spanShort"] = tooShort
if r1.tname.endswith('e5'):
stats["seed1Trim"] = r1.tstart
logging.debug('trim1 %d' % (r1.tstart))
else:
stats["seed1Trim"] = r1.tseqlength - r1.tend
logging.debug('trim1else %d' % (r1.tseqlength - r1.tend))
if r2.tname.endswith('e5'):
stats["seed2Trim"] = r2.tstart
logging.debug('trim2 %d' % (r2.tstart))
else:
stats["seed2Trim"] = r2.tseqlength - r2.tend
logging.debug('trimelse %d' % (r2.tseqlength - r2.tend))
c = singleExtendLookup(a, r1)
if c is not None:
bestF1E = c
c = singleExtendLookup(b, r2)
if c is not None:
bestF2E = c
#sfout.close()
#sfout = sfout.name
#f1fout.close()
#f1fout = f1fout.name
#f2fout.close()
#f2fout = f2fout.name
logging.info("%d reads span" % stats["spanCount"])
logging.info("%d reads extend flank 1" % stats["extendF1Count"])
logging.info("%d reads extend flank 2" % stats["extendF2Count"])
#nt = namedtuple("SubInfo", "stats spanReads flank1Reads flank2Reads spanSeed flank1Seed flank2Seed")
nt = namedtuple("SubInfo", "stats spanSeed flank1Seed flank2Seed")
#seeds out files
ssfout = None
f1sfout = None
f2sfout = None
#replace too short with N's
#if stats["spanCount"] == 0 and len(tooShort) > (stats["extendF1Count"] + stats["extendF2Count"])/2:
"""This is when I would say "oh, i'm too short - and stop early. Now, I'm still going to try to write the
short stuff and treat it like anything else. It'll be up to later processes to catch this guy.
if stats["spanCount"] != 0 and stats["spanShort"]:
#stats["avgSpanBases"] =
#stats["spanCount"] = len(tooShort)
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
logging.debug("but I'm too short")
#stats["fillSeq"] = "N"* abs(stats["spanSeedStart"] - stats["spanSeedEnd"])
stats["fillSeq"] = tooShortSeq
stats["spanSeedScore"] = -500
stats["spanSeedStrand1"] = '0'
stats["spanSeedStrand2"] = '0'
#stats["spanSeedName"] = "tooShortNs"
#ret = nt(stats, None, None, None, None, None, None)
ret = nt(stats, None, None, None)
return ret
"""
if stats["spanCount"] > 0:
stats["avgSpanBases"] = stats["avgSpanBases"] / stats["spanCount"]
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
#write seed
if len(bestSpan.seq) < 50:
logging.warning(
"fill sequence is small (%dbp) can't call consensus" %
(len(bestSpan.seq)))
#I don't know what to return here
ssfout = NamedTemporaryFile(prefix="span_",
suffix=".fasta",
delete=False,
dir=basedir)
ALLTEMPFILES.append(ssfout.name)
logging.debug("spanning with %s" % (bestSpan.name))
ssfout.write(">%s\n%s\n" % (bestSpan.name, bestSpan.seq))
ssfout.close()
ssfout = ssfout.name
#if stats["extendF1Count"] > 0:
if bestF1E is not None:
stats[
"avgExtF1Bases"] = stats["avgExtF1Bases"] / stats["extendF1Count"]
logging.info("estimated flank 1 extend len %d" %
(stats["avgExtF1Bases"]))
#write seed
if len(bestF1E.seq) < 50:
logging.warning(
"f1e sequence is small (%dbp) can't call consensus" %
(len(bestF1E.seq)))
#I don't know what to return here
f1sfout = NamedTemporaryFile(prefix="flank1_",
suffix=".fasta",
delete=False,
dir=basedir)
ALLTEMPFILES.append(f1sfout.name)
f1sfout.write(">%s\n%s\n" % (bestF1E.name, bestF1E.seq))
f1sfout.close()
f1sfout = f1sfout.name
#if stats["extendF2Count"] > 0:
if bestF2E is not None:
stats[
"avgExtF2Bases"] = stats["avgExtF2Bases"] / stats["extendF2Count"]
logging.info("estimated flank 2 extend len %d" %
(stats["avgExtF2Bases"]))
#write seed
if len(bestF2E.seq) < 50:
logging.warning(
"f2e sequence is small (%dbp) can't call consensus" %
(len(bestF2E.seq)))
#I don't know what to return here
f2sfout = NamedTemporaryFile(prefix="flank2",
suffix=".fasta",
delete=False,
dir=basedir)
ALLTEMPFILES.append(f2sfout.name)
f2sfout.write(">%s\n%s\n" % (bestF2E.name, bestF2E.seq))
f2sfout.close()
f2sfout = f2sfout.name
#all of the info I need to return... refactor later and create useful objects
#ret = nt(stats, sfout, f1fout, f2fout, ssfout, f1sfout, f2sfout)
ret = nt(stats, ssfout, f1sfout, f2sfout)
#seeds writing
return ret
import sys
from pbsuite.utils.FileHandlers import FastqFile, M5File
from pbsuite.jelly.Support import AlignmentConnector, SUPPORTFLAGS
"""
Need to do work here
"""
if __name__ == '__main__':
connector = AlignmentConnector()
aligns = connector.parseAlignments(M5File(sys.argv[1]))
reads = FastqFile(sys.argv[2])
bestScore = None
best = None
fout = open("reads.fastq",'w')
spanCount = 0
for readGroup in aligns:
if readGroup[0].qname.startswith("ref"):
continue
if len(readGroup) == 2:
r1, r2 = readGroup
a = connector.extendsTarget(r1)
b = connector.extendsTarget(r2)
if a != SUPPORTFLAGS.none and b != SUPPORTFLAGS.none:
spanCount += 1
print r1.qname, "spans"
rStart = min(r1.qend, r2.qend)
rEnd = max(r1.qstart, r2.qstart)
t = reads[r1.qname].subSeq(rStart, rEnd)
def preunitereads(inputFastq, args):
"""
sent query, I'm going to pop all of the united reads onto this
"""
global ALLTEMPFILES
alignFile = NamedTemporaryFile(prefix="uni_", suffix=".m5", delete=False, dir=args.tempDir).name
ALLTEMPFILES.append(alignFile)
readFile = NamedTemporaryFile(prefix="uni_", suffix=".fasta", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(readFile.name)
input = FastqFile(inputFastq)
for read in input:
readFile.write(">%s\n%s\n" % (input[read].name, input[read].seq))
readFile.close()
readFile = readFile.name
blasr(readFile, readFile, bestn=5, nCandidates=20, nproc=args.nproc, outname=alignFile)
aligns = M5File(alignFile)
con = AlignmentConnector()
extenders = []
for a in aligns:
if a.tname == a.qname:
continue
if a.qstart - a.qend < 500 or a.tstart - a.tend < 500:
continue
sup = con.extendsTarget(a, minCovers=500)
#sup = con.extendsTarget(a, minCovers=100)
a.support = sup
if sup in [SUPPORTFLAGS.left, SUPPORTFLAGS.right]:
extenders.append(a)
best = {}#best of queries
for i in extenders:
score = 0
if i.qname in best:
score = best[i.qname].score
if i.score < score:
best[i.qname] = i
#print "q"
#for i in best.values():
#print str(i)
best2 = {}#best of targets
for i in best.values():
score = 0
if i.tname in best2:
score = best2[i.tname].score
if i.score < score:
best2[i.tname] = i
#print "t"
#for i in best2.values():
#print str(i)
best3 = {}#best of both
for i in best2.values():
keys = [i.qname, i.tname]
keys.sort()
keys = "".join(keys)
score = 0
if keys in best3:
score = best3[keys].score
if i.score < score:
best3[keys] = i
#print 'b'
#for i in best3.values():
#print str(i)
reads = FastqFile(inputFastq)
fout = open(inputFastq, 'a')
count = 0
for i in best3.values():
qseq = None
if i.support == SUPPORTFLAGS.left:
if i.qstrand == '0':
qseq = reads[i.qname].seq + reads[i.tname].seq[i.tend:]
elif i.qstrand == '1':
qseq = reads[i.qname].seq + reads[i.tname].seq[i.tend:].translate(revComp)
if i.support == SUPPORTFLAGS.right:
if i.qstrand == '0':
qseq = reads[i.tname].seq[:i.tstart] + reads[i.qname].seq
elif i.qstrand == '1':
qseq = reads[i.tname].seq[:i.tstart].translate(revComp) + reads[i.qname].seq
if qseq is not None:
count += 1
fout.write("@%s_%s\n%s\n+\n%s\n" % (i.qname, i.tname, qseq, "!"*len(qseq)))
logging.info("Preunited %d reads" % (count))
fout.close()
def getSubSeqs(alignmentFile, readsFile, sameStrand, seeds, predictedGapSize, maxTrim, maxWiggle, basedir="./"):
"""
Finds the seqs that align to the flanks the best, creates a fastq of supporting reads
and the seed
Might have a problem with my best read no going off the edge fully
so I put the maxFlank at 20
I should do more strand correction here
"""
global ALLTEMPFILES
def singleExtendLookup(sup, a):
"""
For getting how a single read extends a single flank
"""
if sup == SUPPORTFLAGS.none:
return None
#set the coordinates of the extending sequence
logging.debug(sup)
logging.debug(a.qname)
mystart = None
myend = None
if a.tname.endswith("e5") and sup in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
if a.tstrand == '0':
mystart = 0
myend = a.qstart
else:
mystart = a.qend
myend = a.qseqlength
elif a.tname.endswith("e3") and sup in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
if a.tstrand == '0':
mystart = a.qend
myend = a.qseqlength
else:
mystart = 0
myend = a.qstart
if mystart is None or myend is None or mystart < 0 or myend > a.qseqlength:
return None
#tscore = a.score * (myend - mystart)
#what flank and is it the best
if a.tname.replace('/','.') == stats["seed1"]:
stats["extendF1Count"] += 1
stats["avgExtF1Bases"] += a.qstart
stats["support"][1].append( sup )
if a.score < stats["extendF1SeedScore"]:
stats["extendF1SeedScore"] = a.score #tscore
stats["extendF1SeedName"] = a.qname
stats["extendF1SeedStart"] = mystart
stats["extendF1SeedEnd"] = myend
stats["extendF1SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f1fout
elif a.tname.replace('/','.') == stats["seed2"]:
stats["extendF2Count"] += 1
stats["avgExtF2Bases"] += a.qstart
stats["support"][2].append( sup )
if a.score < stats["extendF2SeedScore"]:
stats["extendF2SeedScore"] = a.score#tscore
stats["extendF2SeedName"] = a.qname
stats["extendF2SeedStart"] = mystart
stats["extendF2SeedEnd"] = myend
stats["extendF2SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f2fout
#myOut.write(str(reads[a.qname].subSeq(mystart, myend)))
return None
connector = AlignmentConnector()
#aligns = connector.parseAlignments(M5File(alignmentFile))
#no need to connect with the tailmap
aligns = defaultdict(list)
for a in M4File(alignmentFile):
aligns[a.qname].append(a)
aligns = aligns.values()
reads = FastqFile(readsFile)
stats = createStats()
stats["seed1"], stats["seed2"] = seeds
stats["sameStrand"] = sameStrand
bestSpan = None
bestF1E = None
bestF2E = None
for readGroup in aligns:
if len(readGroup) > 2:
best = 0
worst = 0
keep = []
for i in readGroup:
if i.score < best:
keep.insert(0, i)
if len(keep) >= 2:
keep.pop()
best = i.score
elif i.score < worst:
keep.insert(1,i)
if len(keep) >=2:
keep.pop()
worst = i.score
readGroup = keep
if len(readGroup) == 2:
#make sure that the two hits aren't hitting the same target
if readGroup[0].tname == readGroup[1].tname:
if readGroup[0].score <= readGroup[1].score:
del(readGroup[1])
else:
del(readGroup[0])
#hit on each flank
if len(readGroup) == 2:
r1, r2 = readGroup
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a = connector.extendsTarget(r1, maxFlank=maxTrim, minCovers=0)
logging.debug(a)
#Also check appropriate orientation
if r1.tname.endswith('e3'):
if a not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
elif r1.tname.endswith('e5'):
if a not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
b = connector.extendsTarget(r2, maxFlank=maxTrim, minCovers=0)
if r2.tname.endswith('e3'):
if b not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif r2.tname.endswith('e5'):
if b not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif len(readGroup) == 1:
r1 = readGroup[0]
r2 = None
a = connector.extendsTarget(r1, maxFlank=10)
b = SUPPORTFLAGS.none
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a, b = b, a
else:
logging.warning("read %s gave too many alignments" % (readGroup[0].qname))
#it extends both flanks
if a != SUPPORTFLAGS.none and b != SUPPORTFLAGS.none:
logging.debug("%s spans" % r1.qname)
logging.debug("aflag %d bflag %d" % (a,b))
logging.debug("hit1- %s (%d, %d)" % (r1.tname, r1.qstart, r1.qend))
logging.debug("hit2- %s (%d, %d)" % (r2.tname, r2.qstart, r2.qend))
rStart = min(r1.qend, r2.qend)
rEnd = max(r1.qstart, r2.qstart)
sz = rEnd - rStart
tooShort = False
if sz < 50:
logging.info("fill seq is too short to call consensus")
tooShort = True
tooShortSeq = reads[r1.qname].subSeq(rStart, rEnd)
#continue
if predictedGapSize is not None and (predictedGapSize - sz) > maxWiggle:
logging.info("fill seq size %d is smaller than allowed predicted gap size wiggle %d" % (sz, maxWiggle))
continue
#Need to ensure that it's extending in the correct orientation
#need to ensure that everything is on the right strand
if sameStrand and r1.tstrand != r2.tstrand:
logging.debug("bad strandedness")
continue
#check for negative gaps
stats["spanCount"] += 1
stats["avgSpanBases"] += rEnd - rStart
stats["support"][0].append(SUPPORTFLAGS.span)
t = reads[r1.qname].subSeq(rStart, rEnd)
#sfout.write(str(t))
#is it the best spanner
score = r1.score + r2.score
if score < stats["spanSeedScore"]:
logging.debug("scoring %s %s" % (r1.qname, r2.qname))
stats["spanSeedScore"] = score
spanSeedName = r1.qname
stats["spanSeedStrand1"] = r1.tstrand
bestSpan = reads[r1.qname].subSeq(rStart, rEnd)
stats["spanSeedName"] = r1.qname
stats["spanSeedStart"] = rStart
stats["spanSeedEnd"] = rEnd
stats["spanSeedStrand2"] = r2.tstrand
stats["spanShort"] = tooShort
if r1.tname.endswith('e5'):
stats["seed1Trim"] = r1.tstart
logging.debug('trim1 %d' % (r1.tstart))
else:
stats["seed1Trim"] = r1.tseqlength - r1.tend
logging.debug('trim1else %d' % (r1.tseqlength - r1.tend))
if r2.tname.endswith('e5'):
stats["seed2Trim"] = r2.tstart
logging.debug('trim2 %d' % (r2.tstart))
else:
stats["seed2Trim"] = r2.tseqlength - r2.tend
logging.debug('trimelse %d' % (r2.tseqlength - r2.tend))
c = singleExtendLookup(a, r1)
if c is not None:
bestF1E = c
c = singleExtendLookup(b, r2)
if c is not None:
bestF2E = c
#sfout.close()
#sfout = sfout.name
#f1fout.close()
#f1fout = f1fout.name
#f2fout.close()
#f2fout = f2fout.name
logging.info("%d reads span" % stats["spanCount"])
logging.info("%d reads extend flank 1" % stats["extendF1Count"])
logging.info("%d reads extend flank 2" % stats["extendF2Count"])
#nt = namedtuple("SubInfo", "stats spanReads flank1Reads flank2Reads spanSeed flank1Seed flank2Seed")
nt = namedtuple("SubInfo", "stats spanSeed flank1Seed flank2Seed")
#seeds out files
ssfout = None
f1sfout = None
f2sfout = None
#replace too short with N's
#if stats["spanCount"] == 0 and len(tooShort) > (stats["extendF1Count"] + stats["extendF2Count"])/2:
"""This is when I would say "oh, i'm too short - and stop early. Now, I'm still going to try to write the
short stuff and treat it like anything else. It'll be up to later processes to catch this guy.
if stats["spanCount"] != 0 and stats["spanShort"]:
#stats["avgSpanBases"] =
#stats["spanCount"] = len(tooShort)
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
logging.debug("but I'm too short")
#stats["fillSeq"] = "N"* abs(stats["spanSeedStart"] - stats["spanSeedEnd"])
stats["fillSeq"] = tooShortSeq
stats["spanSeedScore"] = -500
stats["spanSeedStrand1"] = '0'
stats["spanSeedStrand2"] = '0'
#stats["spanSeedName"] = "tooShortNs"
#ret = nt(stats, None, None, None, None, None, None)
ret = nt(stats, None, None, None)
return ret
"""
if stats["spanCount"] > 0:
stats["avgSpanBases"] = stats["avgSpanBases"]/stats["spanCount"]
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
#write seed
if len(bestSpan.seq) < 50:
logging.warning("fill sequence is small (%dbp) can't call consensus" % (len(bestSpan.seq)))
#I don't know what to return here
ssfout = NamedTemporaryFile(prefix="span_", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(ssfout.name)
logging.debug("spanning with %s" % (bestSpan.name))
ssfout.write(">%s\n%s\n" % (bestSpan.name, bestSpan.seq))
ssfout.close()
ssfout = ssfout.name
#if stats["extendF1Count"] > 0:
if bestF1E is not None:
stats["avgExtF1Bases"] = stats["avgExtF1Bases"]/stats["extendF1Count"]
logging.info("estimated flank 1 extend len %d" % (stats["avgExtF1Bases"]))
#write seed
if len(bestF1E.seq) < 50:
logging.warning("f1e sequence is small (%dbp) can't call consensus" % (len(bestF1E.seq)))
#I don't know what to return here
f1sfout = NamedTemporaryFile(prefix="flank1_", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(f1sfout.name)
f1sfout.write(">%s\n%s\n" % (bestF1E.name, bestF1E.seq))
f1sfout.close()
f1sfout = f1sfout.name
#if stats["extendF2Count"] > 0:
if bestF2E is not None:
stats["avgExtF2Bases"] = stats["avgExtF2Bases"]/stats["extendF2Count"]
logging.info("estimated flank 2 extend len %d" % (stats["avgExtF2Bases"]))
#write seed
if len(bestF2E.seq) < 50:
logging.warning("f2e sequence is small (%dbp) can't call consensus" % (len(bestF2E.seq)))
#I don't know what to return here
f2sfout = NamedTemporaryFile(prefix="flank2", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(f2sfout.name)
f2sfout.write(">%s\n%s\n" % (bestF2E.name, bestF2E.seq))
f2sfout.close()
f2sfout = f2sfout.name
#all of the info I need to return... refactor later and create useful objects
#ret = nt(stats, sfout, f1fout, f2fout, ssfout, f1sfout, f2sfout)
ret = nt(stats, ssfout, f1sfout, f2sfout)
#seeds writing
return ret