def prepare_annotation(self):
"""Prepares for overlapping annotations"""
if self.genome:
# for conversion of chromosome name between FASTA and annotation (hg19)
self.chrom_proper = tools.ucsc_chroms(self.genome, self.annodir)
splice_motif_file = os.path.join(self.annodir, self.genome, 'splice_motifs.txt')
if os.path.isfile(splice_motif_file):
self.splice_motif_file = splice_motif_file
# for constructing cDNA sequences for in/out frame determination
self.refseq = tools.get_refseq_from_2bit(self.annodir, self.genome)
# for finding genes, exons, etc
if self.gene_model:
self.ff = FeatureFinder(self.genome, self.gene_model, self.annodir, self.mmcfg)
# for getting rid of contigs that mapped entirely with repeats
self.repeat_overlaps = repeat.prepare_overlap(self.genome, self.annodir)
class SNVCaller:
"""Calls indels and SNVs from single, non-split contig alignment"""
event_types = ('snv', 'ins', 'del', 'inv', 'dup', 'ITD')
output_headers = ['id',
'type',
'chr',
'chr_start',
'chr_end',
'ctg',
'ctg_len',
'ctg_start',
'ctg_end',
'len',
'ref',
'alt',
'event_reads',
'contig_reads',
'genome_reads',
'gene',
'repeat-length',
'ctg_strand',
'from_end',
'confirm_contig_region',
'within_simple_repeats',
'repeatmasker',
'within_segdup',
'at_least_1_read_opposite',
'dbsnp'
]
def __init__(self, genome, get_snv, get_indel, align_file=None, contigs_file=None, min_reads=0,
debug=False, min_from_end=None, bubble_mapping=None, sample_type='transcriptome', gene_model=None, annodir=None, mmcfg=None):
self.align_file = align_file
self.contigs_file = contigs_file
self.genome = genome
self.min_reads = min_reads
self.debug = debug
self.min_from_end = min_from_end
self.gene_model = gene_model
self.annodir = annodir
self.mmcfg = mmcfg
self.chrom_proper, self.refseq, self.ff, self.repeat_overlaps, self.splice_motif_file = None, None, None, None, None
if self.genome:
self.prepare_annotation()
self.snvs = []
self.grouped_snvs = {}
self.bubble_mapping = bubble_mapping
self.sample_type = sample_type
# actions to perform
self.get_snv = get_snv
self.get_indel = get_indel
def prepare_annotation(self):
"""Prepares for overlapping annotations"""
if self.genome:
# for conversion of chromosome name between FASTA and annotation (hg19)
self.chrom_proper = tools.ucsc_chroms(self.genome, self.annodir)
splice_motif_file = os.path.join(self.annodir, self.genome, 'splice_motifs.txt')
if os.path.isfile(splice_motif_file):
self.splice_motif_file = splice_motif_file
# for constructing cDNA sequences for in/out frame determination
self.refseq = tools.get_refseq_from_2bit(self.annodir, self.genome)
# for finding genes, exons, etc
if self.gene_model:
self.ff = FeatureFinder(self.genome, self.gene_model, self.annodir, self.mmcfg)
# for getting rid of contigs that mapped entirely with repeats
self.repeat_overlaps = repeat.prepare_overlap(self.genome, self.annodir)
def extract(self, cutoff=None, no_group=False, match_percent=None, identity=None, no_segdup=False):
"""Wrapper function for identifying indels and SNVs from non-split alignments"""
splice_motifs = tools.get_splice_motifs(self.splice_motif_file)
filters = {'unique':True, 'bestn':1, 'match':match_percent, 'identity':identity}
# extracts alignments
out_format = os.path.splitext(self.align_file)[1]
aligns = {
'.psl': psl.parse,
'.sam': sam.parse
}[out_format](self.align_file, filters, splice_motif_file=self.splice_motif_file, refseq=self.refseq, noline=True)
# links contig sequence to alignment
ass = Assembly(None, k=1)
ass.fasta = self.contigs_file
contigs = ass.get_contigs(sequence=True)
contig_dict = dict((contig.num, contig) for contig in contigs)
for align in aligns:
if contig_dict.has_key(align.query):
align.contig = contig_dict[align.query]
for align in aligns:
if self.bubble_mapping and not self.bubble_mapping.is_bubble_mapped_to_contig(align.query):
print "remove bubble", align.query
continue
snvs = self.get_snvs(align, splice_motifs=splice_motifs, cutoff=cutoff, no_segdup=no_segdup)
for snv in snvs:
snv.var_len = align.query_len
snv.from_end = min(int(snv.var_start) - int(align.qstart), int(align.qend) - int(snv.var_end))
# identifies repeat units
snv.upshift(self.refseq)
snv.expand_contig_region(align.contig.sequence, align.query_strand)
target = snv.ref
# re-labels 'ins' as 'dup' if expansion >=2 and length > 3
if snv.snv_type == 'ins' and snv.expansion >= 2 and snv.snv_len > 3:
snv.snv_type = 'dup'
snv.ref_start += 1
snv.ref_end = snv.ref_start + snv.snv_len - 1
self.snvs.append(snv)
# group events
if not no_group:
self.grouped_snvs = self.group(self.snvs)
def annotate_genes(self):
"""Annotates events with genes"""
if self.ff:
for groups in self.grouped_snvs.values():
for snvs in groups.values():
gene = self.ff.get_feature(' '.join((tools.proper_chrom(snvs[0].ref,
chrom_proper=self.chrom_proper),
str(snvs[0].ref_start),
str(snvs[0].ref_end))),
refseq=self.refseq,
variant=snvs[0].var_seq,
change=snvs[0].snv_type,
chrom=snvs[0].ref)
for snv in snvs:
snv.gene = gene
# relabel duplication within exon as ITD
if snvs[0].snv_type == 'dup' and 'exon' in snvs[0].gene\
and not 'utr' in snvs[0].gene and not 'intron' in snvs[0].gene:
for snv in snvs:
snv.snv_type = 'ITD'
def is_within_repeats(self, proper_chrom, span):
"""Determines if given coordinate span overlaps segdups or simple_repeats"""
overlaps = repeat.find_overlaps({'chrom':proper_chrom, 'start':span[0], 'end':span[1]}, self.repeat_overlaps)
if overlaps:
for repeat_type in overlaps.keys():
if overlaps[repeat_type]:
if repeat_type == 'segdup' or repeat_type == 'simple_repeats':
return True
return False
def overlap_repeats(self):
"""Overlaps event coordinates with repeatmasker, simple repeats"""
event_groups_by_chr = self.group_by_chr()
for chrom, events in event_groups_by_chr.iteritems():
proper_chrom = tools.proper_chrom(chrom, chrom_proper=self.chrom_proper)
for snv_type, snv_groups in events.iteritems():
print 'processing repeat', snv_type
for snvs in snv_groups:
overlaps = repeat.find_overlaps({'chrom':proper_chrom, 'start':int(snvs[0].ref_start),
'end':int(snvs[0].ref_end)}, self.repeat_overlaps)
if overlaps:
attrs = {}
for repeat_type, types in overlaps.iteritems():
if repeat_type == 'simple_repeats':
attr = 'within_simple_repeats'
elif repeat_type == 'segdup':
attr = 'within_segdup'
elif repeat_type == 'rmsk':
attr = 'repeatmasker'
if types:
# only report one with shortest name
types_sorted = types.keys()
types_sorted.sort(lambda x,y: len(x)-len(y))
attrs[attr] = types_sorted[0]
if attrs:
for snv in snvs:
tools.set_attrs(snv, attrs)
# clears cache
for repeat_olap in self.repeat_overlaps.values():
repeat_olap.finish()
def group_by_chr(self):
"""Groups events by genomic location for overlapping with annotation such as dbSNP"""
events_by_chr = {}
for event_type in self.event_types:
if not self.grouped_snvs.has_key(event_type):
continue
for key, snvs in self.grouped_snvs[event_type].iteritems():
if not events_by_chr.has_key(snvs[0].ref):
events_by_chr[snvs[0].ref] = {}
events_by_chr[snvs[0].ref] = {event_type:[]}
if not events_by_chr[snvs[0].ref].has_key(event_type):
events_by_chr[snvs[0].ref][event_type] = []
events_by_chr[snvs[0].ref][event_type].append(snvs)
return events_by_chr
def overlap_dbsnp(self):
"""Overlaps events with dbSNP"""
event_groups_by_chr = self.group_by_chr()
for chrom, events in event_groups_by_chr.iteritems():
proper_chrom = tools.proper_chrom(chrom, chrom_proper=self.chrom_proper)
snp_overlap = dbsnp.prepare_overlap(self.genome, proper_chrom, self.annodir)
for snv_type, snv_groups in events.iteritems():
event_type_check = snv_type
if snv_type in ('dup', 'ITD', 'PTD'):
event_type_check = 'ins'
for snvs in snv_groups:
start, end = int(snvs[0].ref_start), int(snvs[0].ref_end)
if snv_type in ('dup', 'ITD', 'PTD'):
start, end = int(snvs[0].ref_start) - 1, int(snvs[0].ref_start) - 1
known = dbsnp.find_concordants({'type':event_type_check, 'chrom':proper_chrom, 'start':start, 'end':end, 'allele':snvs[0].var_seq.lower(), 'size':int(snvs[0].snv_len)}, snp_overlap, refseq=self.refseq, target=chrom)
if known:
for snv in snvs:
snv.dbsnp = ','.join(known)
snp_overlap.finish()
def compare_contig(self, snv1, snv2):
"""For sorting events by comparing contig number and coordinates"""
if snv1.var.isdigit() and snv2.var.isdigit():
if int(snv1.var) < int(snv2.var):
return -1
elif int(snv1.var) > int(snv2.var):
return 1
elif int(snv1.var_start) < int(snv2.var_start):
return -1
elif int(snv1.var_start) > int(snv2.var_start):
return 1
else:
return 0
elif snv1.var.isdigit() and not snv2.var.isdigit():
return -1
elif not snv1.var.isdigit() and snv2.var.isdigit():
return 1
else:
if snv1.var < snv2.var:
return -1
elif snv1.var > snv2.var:
return 1
elif int(snv1.var_start) < int(snv2.var_start):
return -1
elif int(snv1.var_start) > int(snv2.var_start):
return 1
else:
return 0
def add_support(self, lib=None, from_end=None, genome_bamfile=None, contigs_bamfile=None):
"""Calculates read support"""
genome_bam = None
contigs_bam = None
if genome_bamfile and os.path.exists(genome_bamfile):
genome_bam = BAM(genome_bamfile)
for event_type in sorted(self.grouped_snvs.keys()):
for coord_allele in self.grouped_snvs[event_type]:
snvs = self.grouped_snvs[event_type][coord_allele]
snvs[0].genome_read_support(genome_bam, self.refseq, from_end=from_end)
for i in range(1, len(snvs)):
snvs[i].nreads_genome = snvs[0].nreads_genome
if contigs_bamfile and os.path.exists(contigs_bamfile):
contigs_bam = BAM(contigs_bamfile, min_mapq=0)
self.snvs.sort(self.compare_contig)
for event in self.snvs:
event.contig_read_support(contigs_bam, lib=lib, get_reads=True, from_end=from_end)
def group(self, snvs):
"""Groups events by event type, coordinate, and allele sequence"""
grouped_snvs = {}
for snv in snvs:
if snv.artefact:
continue
seq = ""
if snv.snv_type in self.event_types:
seq = snv.var_seq[:]
coord = snv.coord()
snv_key = coord + "-" + seq
if not grouped_snvs.has_key(snv.snv_type):
grouped_snvs[snv.snv_type] = {snv_key:[snv]}
else:
if not grouped_snvs[snv.snv_type].has_key(snv_key):
grouped_snvs[snv.snv_type][snv_key] = [snv]
else:
grouped_snvs[snv.snv_type][snv_key].append(snv)
return grouped_snvs
def add_contig_reads(self, grouped_snvs):
"""Sums contig read supports of members within same group"""
for type in self.event_types:
if not grouped_snvs.has_key(type):
continue
for coord in grouped_snvs[type].keys():
event_reads = 0
for snv in grouped_snvs[type][coord]:
if snv.nreads_contig != 'na':
event_reads += int(snv.nreads_contig)
for snv in grouped_snvs[type][coord]:
snv.nreads_event = event_reads
def output_groups(self, grouped_snvs, output, debug=False):
"""Outputs grouped events with identifiers indicating groups"""
# group count
count1 = 1
for type in self.event_types:
if not grouped_snvs.has_key(type):
continue
coords = sorted(grouped_snvs[type].keys(), key=lambda coord_allele: (grouped_snvs[type][coord_allele][0].ref, int(grouped_snvs[type][coord_allele][0].ref_start)))
for coord in coords:
# group-member count
count2 = 1
for snv in grouped_snvs[type][coord]:
if len(grouped_snvs[type][coord]) == 1:
count = count1
else:
count = "%d.%d" % (count1, count2)
output.write(str(count) + "\t" + snv.tab(debug=debug))
count2 += 1
count1 += 1
def report(self, outdir, post_filter=False):
"""Produces different output files"""
txt_file = outdir + "/events.tsv"
self.output_txt(self.snvs, txt_file)
if post_filter:
self.post_filter(min_from_end=self.min_from_end)
if self.sample_type == 'genome':
filtered_snvs = [snv for snv in self.snvs if not snv.artefact and snv.enough_coverage and not snv.too_close_to_end and snv.at_least_1_read_opposite ]
else:
filtered_snvs = [snv for snv in self.snvs if not snv.artefact and snv.enough_coverage and not snv.too_close_to_end ]
txt_file = outdir + "/events_filtered.tsv"
self.output_txt(filtered_snvs, txt_file)
exon_snvs = [snv for snv in filtered_snvs if snv.exon and snv.nonsynon]
txt_file = outdir + "/events_exons.tsv"
self.output_txt(exon_snvs, txt_file)
debug_out = open(outdir + "/filter_debug.tsv", 'w')
self.output_groups(self.grouped_snvs, debug_out, debug=True)
debug_out.close()
novel_snvs = [snv for snv in filtered_snvs if snv.dbsnp == '-']
txt_file = outdir + "/events_filtered_novel.tsv"
self.output_txt(novel_snvs, txt_file)
novel_exon_snvs = [snv for snv in filtered_snvs if snv.exon and snv.nonsynon and snv.dbsnp == '-']
txt_file = outdir + "/events_exons_novel.tsv"
self.output_txt(novel_exon_snvs, txt_file)
def output_txt(self, snvs, outfile):
"""Groups and outputs given events in tabular format"""
grouped_snvs = self.group(snvs)
out = open(outfile, 'w')
out.write("\t".join(self.output_headers) + "\n")
self.output_groups(grouped_snvs, out)
out.close()
def post_filter(self, min_from_end=None, no_mito=False):
"""Filters events based on read-support, mitonchondria, distance from contig edge, etc"""
snvs_filtered = []
# for extracting kmer length from contig name
kmer = re.compile(r'k(\d+):')
# filters out SNVs that are highly clustered
self.filter_bad_region()
for snv in self.snvs:
if snv.nreads_event != 'na' and int(snv.nreads_event) >= int(self.min_reads):
snv.enough_coverage = True
if snv.nreads_genome != 'na' and int(snv.nreads_genome) < int(self.min_reads_genome):
snv.enough_coverage = False
if snv.gene and ('exon' in snv.gene or 'utr' in snv.gene):
snv.exon = True
if snv.exon and not ':synon' in snv.gene:
snv.nonsynon = True
# min_from_end only applies for non-bubbles (contig name ended with \d)
if re.match('\d', snv.var[-1]):
if min_from_end == None:
m = kmer.search(snv.var)
if m and m.group:
min_from_end_contig = int(m.group(1))
if int(snv.from_end) < min_from_end_contig:
snv.too_close_to_end = True
elif int(snv.from_end) < int(min_from_end):
snv.too_close_to_end = True
# mitochondria
if no_mito and re.match('m', snv.ref, re.IGNORECASE):
snv.artefact = True
def filter_bad_region(self):
"""Filters out SNVs that are highly clustered.
Cannot have more than 10 in 500bp windown.
"""
# group events by contig
snvs_by_contig = {}
for snv in self.snvs:
if snv.snv_type != 'snv' or len(snv.var_seq) != 1:
continue
if not snvs_by_contig.has_key(snv.var):
snvs_by_contig[snv.var] = []
snvs_by_contig[snv.var].append(snv)
for contig, snvs in snvs_by_contig.iteritems():
snvs.sort(lambda x,y: int(x.var_start)-int(y.var_start))
for i in range(len(snvs)):
if i < len(snvs)-1:
diff = int(snvs[i+1].var_start) - int(snvs[i].var_start)
else:
diff = 'na'
max_in_window = 10
window = 500
for i in range(len(snvs)-max_in_window+1):
bad = [i]
for j in range(i+1, len(snvs)):
if int(snvs[j].var_start) - int(snvs[i].var_start) < window:
bad.append(j)
else:
break
if len(bad) > max_in_window:
for idx in bad:
snvs[idx].artefact = True
def get_snvs(self, align, splice_motifs=None, cutoff=None, no_segdup=False):
"""Extracts indels and snvs from alignment"""
all_snvs = []
sys.stderr.write("processing %s\n" % (align.query))
if self.refseq:
if self.sample_type == 'transcriptome' and self.fix_align:
align.correct_blocks(splice_motifs, self.refseq, align.contig.sequence)
if self.get_indel:
all_snvs.extend(self.gap_snv(align, splice_motifs, align.contig.sequence, cutoff=cutoff))
if self.get_snv and (align.mismatch is None or int(align.mismatch) > 0):
all_snvs.extend(self.match_blocks(align, self.refseq, align.contig.sequence))
if all_snvs and no_segdup:
proper_chrom = tools.proper_chrom(align.target, chrom_proper=self.chrom_proper)
if self.is_within_repeats(proper_chrom, [int(align.tstart), int(align.tend)]):
print 'skip contig %s (%sbp): %s:%s-%s entirely with segdup/repeat' % (align.query, align.query_len, align.target, align.tstart, align.tend)
del all_snvs[:]
return all_snvs
def match_intron(self, ss, splice_motifs):
"""Determines splite sites correspond to intron by comparing to splice motifs"""
if ss and (splice_motifs.has_key(ss.lower()) or splice_motifs.has_key(tools.reverse_complement(ss).lower())):
return True
else:
return False
def gap_snv(self, align, splice_motifs, query_seq, cutoff=None):
"""Identifies insertions, deletions, inversion from gapped alignments"""
if self.debug:
print align.target, align.blocks
print align.query, align.query_blocks
print align.splice_sites
snvs = []
# cannot identify indels without splice site information
if self.sample_type == 'transcriptome' and not align.splice_sites:
return snvs
for i in range(len(align.blocks)-1):
if self.sample_type != 'transcriptome' or not self.match_intron(align.splice_sites[i], splice_motifs):
if align.query_strand == '+':
qstart = align.query_blocks[i][1]+1
qend = align.query_blocks[i+1][0]-1
query = query_seq[qstart-1:qend]
else:
qend = align.query_blocks[i][1]-1
qstart = align.query_blocks[i+1][0]+1
query = query_seq[qstart-1:qend]
query = tools.reverse_complement(query)
# target strand always + from psl
tstart = align.blocks[i][1]+1
tend = align.blocks[i+1][0]-1
target = ''
if tstart <= tend:
target = self.refseq.GetSequence(align.target, tstart, tend)
#if code cannot extract sequence from reference, there must be a disagreement between alignment and reference - abort analysis
if tend > tstart-1 and len(target) < 1:
sys.stderr.write("cannot extract reference sequence, abort: %s %s %s\n" % (align.target, tstart-1, tend))
sys.exit(100)
snv_type = None
if qstart > qend and (tend - tstart) >= 0:
size = tend - tstart + 1
if align.query_strand == '+':
qstart = qend
else:
qend = qstart
snv_type = "del"
elif tstart > tend and (qend - qstart) >= 0:
size = qend - qstart + 1
tstart = tend
snv_type = "ins"
else:
size = min(1, tend - tstart + 1)
snv_type = "indel"
# skip if 0 or negative size event detected (or smaller than cutoff)
if size <= 0 or (cutoff and size > cutoff):
continue
target = target.lower()
query = query.lower()
# would not report event with non-AGCT characters
if not re.search('[^agtcATGC]', target) and not re.search('[^agtcATGC]', query):
if snv_type != 'indel':
snv = SNV('psl', snv_type, align.target, tstart, tend, target, align.query_strand, align.query, qstart, qend, query)
snvs.append(snv)
# resolves indels
else:
if len(query) == len(target) and\
(query[::-1].lower() == target.lower() or tools.reverse_complement(query).lower() == target.lower()):
# inversion must be longer than 1 base
if len(query) > 1:
snv = SNV('psl', 'inv', align.target, tstart, tend, target, align.query_strand, align.query, qstart, qend, query)
snvs.append(snv)
# 1 bp gap in both query and target == snv
else:
snv = SNV('psl', 'snv', align.target, tstart, tend, target, align.query_strand, align.query, qstart, qend, query)
snvs.append(snv)
# breaks up indel into ins and del
else:
if align.query_strand == '+':
qcoord = qstart
else:
qcoord = qend
snv = SNV('psl', 'del', align.target, tstart, tend, target, align.query_strand, align.query, qcoord, qcoord, query)
snvs.append(snv)
tcoord = tstart
snv = SNV('psl', 'ins', align.target, tcoord, tcoord, target, align.query_strand, align.query, qstart, qend, query)
snvs.append(snv)
return snvs
def parse_results(self, snv_file, select_types=None, chrom=None):
"""Parses results from single file into SNV objects"""
names = SNVCaller.output_headers
# conversion between header name and object attribute
field_name_conversion = {
'type': 'snv_type',
'chr': 'ref',
'chr_start': 'ref_start',
'chr_end': 'ref_end',
'ctg': 'var',
'ctg_len': 'var_len',
'ctg_start': 'var_start',
'ctg_end': 'var_end',
'len': 'snv_len',
'ref': 'ref_seq',
'alt': 'var_seq',
'event_reads': 'nreads_event',
'contig_reads': 'nreads_contig',
'genome_reads': 'nreads_genome',
'gene': 'gene',
'from_end': 'from_end',
'ctg_strand': 'query_strand',
}
for line in open(snv_file, 'r'):
cols = line.rstrip('\n').split('\t')
if cols[0] == 'id':
continue
attributes = {}
for i in range(1, len(cols)):
name = names[i]
value = cols[i]
if field_name_conversion.has_key(name):
name = field_name_conversion[name]
if name in ('expansion', 'from_end'):
value = int(value)
elif name == 'confirm_contig_region':
value = value.split('-')
value[0] = int(value[0])
value[1] = int(value[1])
elif name == 'at_least_1_read_opposite':
if value == 'true':
value = True
else:
value = False
attributes[name] = value
if select_types and not attributes['snv_type'] in select_types:
continue
if chrom and attributes['ref'] != chrom:
continue
snv = SNV(method='psl')
tools.set_attrs(snv, attributes)
self.snvs.append(snv)
def parse_results_dir(self, path):
"""Parses results into single file given a directory of output directories"""
output_dirs = sorted(glob.glob(os.path.join(path, '*')))
output_files = []
num_output_dirs = 0
missing_dirs = []
ok = True
for job_num in range(1, len(output_dirs)+1):
cluster_outdir = "%s/%s" % (options.output_file, job_num)
if os.path.isdir(cluster_outdir):
num_output_dirs += 1
snv_file = cluster_outdir + '/events.tsv'
if os.path.exists(snv_file):
output_files.append(snv_file)
else:
missing_dirs.append(str(job_num))
print snv_file
sys.stdout.write("%s does not have output\n" % (cluster_outdir))
ok = False
sys.stdout.write("output dirs:%s output files:%s\n" % (num_output_dirs, len(output_files)))
if num_output_dirs == len(output_files):
concat_outfile = options.outdir + "/events_concat.tsv"
print concat_outfile
concat_out = open(concat_outfile, 'w')
# just for check if header is to be written
count = 0
for output_file in output_files:
infile = open(output_file, 'r')
if count == 0:
concat_out.writelines(open(output_file, 'r').readlines())
else:
concat_out.writelines(open(output_file, 'r').readlines()[1:])
infile.close()
self.parse_results(output_file)
count += 1
concat_out.close()
else:
sys.stdout.write("missing (%s):%s\n" % (len(missing_dirs), ','.join(missing_dirs)))
ok = False
return ok
def match_blocks(self, align, query_seq):
"""Identifies SNVs"""
snvs = []
for i in range(len(align.blocks)):
if align.query_strand == '+':
qseq = query_seq[int(align.query_blocks[i][0])-1:int(align.query_blocks[i][1])]
else:
qseq = tools.reverse_complement(query_seq[int(align.query_blocks[i][1]-1):int(align.query_blocks[i][0])])
tseq = self.refseq.GetSequence(align.target, int(align.blocks[i][0]), int(align.blocks[i][1]))
mismatches = self.find_mismatches(qseq, tseq)
for pos, change in mismatches.iteritems():
tpos = int(align.blocks[i][0]) + pos
if int(align.query_blocks[i][0]) < int(align.query_blocks[i][1]):
qpos = int(align.query_blocks[i][0]) + pos
else:
qpos = int(align.query_blocks[i][0]) - pos
snv = SNV('psl', 'snv', align.target, tpos, tpos, change[0], align.query_strand, align.query, qpos, qpos, change[1])
snvs.append(snv)
return snvs
def find_mismatches(self, qseq, tseq):
"""Reports substitutions given query and target sequence of same length"""
pos = {}
bases = ['a','g','t','c']
if len(qseq) == len(tseq):
for i in range(len(qseq)):
if qseq[i].lower() != tseq[i].lower() and qseq[i].lower() in bases and tseq[i].lower() in bases:
pos[i] = [tseq[i].lower(), qseq[i].lower()]
return pos