def main():
#Parse the inputs args/options
parser = OptionParser(usage="usage: inputFastaFile outputFastaFile outputMutationsFile [options]",
version="%prog 0.1")
parser.add_option("--snpRate", dest="snpRate",
help="The probability of introducing a random different base at each position",
default=0.2, type=float)
#Parse the options/arguments
options, args = parser.parse_args()
#Print help message if no input
if len(sys.argv) == 1:
parser.print_help()
sys.exit(0)
#Exit if the arguments are not what we expect
if len(args) != 3:
raise RuntimeError("Expected three arguments, got: %s" % " ".join(args))
#This call gets the mutated sequences and a list of mutations
mutatedSequences, allMutations = mutateSequences(getFastaDictionary(args[0]), options.snpRate)
#Write out the mutated sequences into the given file
fH = open(args[1], 'w')
for name in mutatedSequences:
fastaWrite(fH, name, mutatedSequences[name])
fH.close()
#Write out mutations
fH = open(args[2], 'w')
for mutation in allMutations:
fH.write("\t".join(map(str, mutation)) + "\n")
fH.close()
def Substitutions(readFastqFile,
referenceFastaFile,
samFile,
outputDir,
kmer=6):
"""Calculates stats on substitutions
"""
refSequences = getFastaDictionary(
referenceFastaFile) #Hash of names to sequences
readSequences = getFastqDictionary(
readFastqFile) #Hash of names to sequences
sM = SubstitutionMatrix() #The thing to store the counts in
sam = pysam.Samfile(samFile, "r")
for aR in samIterator(sam): #Iterate on the sam lines
for aP in AlignedPair.iterator(aR, refSequences[sam.getrname(
aR.rname)], readSequences[
aR.qname]): #Walk through the matches mismatches:
sM.addAlignedPair(aP.getRefBase(), aP.getReadBase())
sam.close()
#Write out the substitution info
open(os.path.join(outputDir, "substitutions.xml"),
'w').write(prettyXml(sM.getXML()))
bases = "ACGT"
outf = open(os.path.join(outputDir, "subst.tsv"), "w")
outf.write("A\tC\tG\tT\n")
for x in bases:
freqs = sM.getFreqs(x, bases)
outf.write("{}\t{}\n".format(x, "\t".join(map(str, freqs)), "\n"))
outf.close()
analysis = str(samFile.split("/")[-1].split(".sam")[0])
system("Rscript scripts/substitution_plot.R {} {} {}".format(
os.path.join(outputDir, "subst.tsv"),
os.path.join(outputDir, "substitution_plot.pdf"), analysis))
def validateVcf(self, vcfFile, referenceFastaFile, mutationsFile):
#Load reference sequences
referenceSequences = getFastaDictionary(referenceFastaFile)
#Load mutations
mutations = set(map(lambda x : (x[0], int(x[1])+1, x[2]), \
map(lambda x : x.split(), open(mutationsFile, 'r'))))
#Load VCF mutations
imputedMutations = vcfRead(vcfFile)
#print "Known mutations", sorted(list(mutations))
#print "Imputed mutations", sorted(list(imputedMutations))
#Compare mutation sets
intersectionSize = float(len(mutations.intersection(imputedMutations)))
#Return precision, recall, number of mutations called, number of known mutations
return intersectionSize/len(imputedMutations) if len(imputedMutations) else 0.0, \
intersectionSize/len(mutations) if len(mutations) else 0.0, len(imputedMutations), len(mutations)
def validateVcf(self, vcfFile, referenceFastaFile, mutationsFile):
#Load reference sequences
referenceSequences = getFastaDictionary(referenceFastaFile)
#Load mutations
mutations = set(map(lambda x : (x[0], int(x[1])+1, x[2]), \
map(lambda x : x.split(), open(mutationsFile, 'r'))))
#Load VCF mutations
imputedMutations = vcfRead(vcfFile)
#print "Known mutations", sorted(list(mutations))
#print "Imputed mutations", sorted(list(imputedMutations))
#Compare mutation sets
intersectionSize = float(len(mutations.intersection(imputedMutations)))
#Return precision, recall, number of mutations called, number of known mutations
return intersectionSize/len(imputedMutations) if len(imputedMutations) else 0.0, \
intersectionSize/len(mutations) if len(mutations) else 0.0, len(imputedMutations), len(mutations)
def main():
#Parse the inputs args/options
parser = OptionParser(
usage=
"usage: inputFastaFile outputFastaFile outputMutationsFile [options]",
version="%prog 0.1")
parser.add_option(
"--snpRate",
dest="snpRate",
help=
"The probability of introducing a random different base at each position",
default=0.2,
type=float)
#Parse the options/arguments
options, args = parser.parse_args()
#Print help message if no input
if len(sys.argv) == 1:
parser.print_help()
sys.exit(0)
#Exit if the arguments are not what we expect
if len(args) != 3:
raise RuntimeError("Expected three arguments, got: %s" %
" ".join(args))
#This call gets the mutated sequences and a list of mutations
mutatedSequences, allMutations = mutateSequences(
getFastaDictionary(args[0]), options.snpRate)
#Write out the mutated sequences into the given file
fH = open(args[1], 'w')
for name in mutatedSequences:
fastaWrite(fH, name, mutatedSequences[name])
fH.close()
#Write out mutations
fH = open(args[2], 'w')
for mutation in allMutations:
fH.write("\t".join(map(str, mutation)) + "\n")
fH.close()
def check_alignments(self,
true_alignments,
reads,
reference,
kmer_length,
contig_name,
extra_args=None):
def get_kmer(start):
return referece_sequence[start:start + kmer_length]
assert len(
glob.glob(reads +
"*.fast5")) > 0, "Didn't find zymo test MinION reads"
assert os.path.isfile(reference), "Didn't find zymo reference sequence"
alignment_command = "./runSignalAlign -d={reads} -r={ref} -smt=threeState -o={testDir} " \
"".format(reads=reads, ref=reference, testDir="./signalAlign_unittest/")
if extra_args is not None:
alignment_command += extra_args
null_output = open(os.devnull, 'w')
result = call(alignment_command,
shell=True,
bufsize=-1,
stdout=null_output,
stderr=null_output)
self.assertTrue(
result == 0, "error running signalAlign alignments command was {}"
"".format(alignment_command))
test_alignments = glob.glob(
"./signalAlign_unittest/tempFiles_alignment/*.tsv")
referece_sequence = getFastaDictionary(reference)[contig_name]
self.assertTrue(
len(test_alignments) == len(glob.glob(true_alignments + "*.tsv")),
"Didn't make all alignments got {got} should be {should}".format(
got=len(test_alignments),
should=len(glob.glob(true_alignments + "*.tsv"))))
for alignment in test_alignments:
alignment_file = alignment.split("/")[-1]
expected = parse_alignment_full(true_alignments + alignment_file)
obs = parse_alignment_full(alignment)
self.assertTrue(len(obs) == len(expected))
for row in obs.itertuples():
ref_pos = row[1]
obs_kmer = row[2]
strand = row[3]
exp_kmer = get_kmer(ref_pos)
self.assertTrue(
obs_kmer == exp_kmer,
msg=
"kmer at index {idx} on strand {strand} is {obs} should be "
"{exp}, file {f}".format(idx=ref_pos,
strand=strand,
obs=obs_kmer,
exp=exp_kmer,
f=alignment))
