#!/usr/bin/env python #Guruprasad Ananda #MAQ mapper for SOLiD colourspace-reads import sys, os, zipfile, tempfile, subprocess def stop_err( msg ): sys.stderr.write( "%s\n" % msg ) sys.exit() def __main__(): out_fname = sys.argv[1].strip() out_f2 = open(sys.argv[2].strip(),'r+') ref_fname = sys.argv[3].strip() f3_read_fname = sys.argv[4].strip() f3_qual_fname = sys.argv[5].strip() paired = sys.argv[6] if paired == 'yes': r3_read_fname = sys.argv[7].strip() r3_qual_fname = sys.argv[8].strip() min_mapqual = int(sys.argv[9].strip()) max_mismatch = int(sys.argv[10].strip()) out_f3name = sys.argv[11].strip() subprocess_dict = {} ref_csfa = tempfile.NamedTemporaryFile() ref_bfa = tempfile.NamedTemporaryFile() ref_csbfa = tempfile.NamedTemporaryFile() cmd2_1 = 'maq fasta2csfa %s > %s 2>&1' %(ref_fname,ref_csfa.name) cmd2_2 = 'maq fasta2bfa %s %s 2>&1' %(ref_csfa.name,ref_csbfa.name) cmd2_3 = 'maq fasta2bfa %s %s 2>&1' %(ref_fname,ref_bfa.name) try: os.system(cmd2_1) os.system(cmd2_2) os.system(cmd2_3) except Exception, erf: stop_err(str(erf)+"Error processing reference sequence") if paired == 'yes': #paired end reads tmpf = tempfile.NamedTemporaryFile() #forward reads tmpr = tempfile.NamedTemporaryFile() #reverse reads tmps = tempfile.NamedTemporaryFile() #single reads tmpffastq = tempfile.NamedTemporaryFile() tmprfastq = tempfile.NamedTemporaryFile() tmpsfastq = tempfile.NamedTemporaryFile() cmd1 = "solid2fastq_modified.pl 'yes' %s %s %s %s %s %s %s 2>&1" %(tmpf.name,tmpr.name,tmps.name,f3_read_fname,f3_qual_fname,r3_read_fname,r3_qual_fname) try: os.system(cmd1) os.system('gunzip -c %s >> %s' %(tmpf.name,tmpffastq.name)) os.system('gunzip -c %s >> %s' %(tmpr.name,tmprfastq.name)) os.system('gunzip -c %s >> %s' %(tmps.name,tmpsfastq.name)) except Exception, eq: stop_err("Error converting data to fastq format." + str(eq)) #make a temp directory where the split fastq files will be stored try: split_dir = tempfile.mkdtemp() split_file_prefix_f = tempfile.mktemp(dir=split_dir) split_file_prefix_r = tempfile.mktemp(dir=split_dir) splitcmd_f = 'split -a 2 -l %d %s %s' %(32000000,tmpffastq.name,split_file_prefix_f) #32M lines correspond to 8M reads splitcmd_r = 'split -a 2 -l %d %s %s' %(32000000,tmprfastq.name,split_file_prefix_r) #32M lines correspond to 8M reads os.system(splitcmd_f) os.system(splitcmd_r) os.chdir(split_dir) ii = 0 for fastq in os.listdir(split_dir): if not fastq.startswith(split_file_prefix_f.split("/")[-1]): continue fastq_r = split_file_prefix_r + fastq.split(split_file_prefix_f.split("/")[-1])[1] #find the reverse strand fastq corresponding to formward strand fastq tmpbfq_f = tempfile.NamedTemporaryFile() tmpbfq_r = tempfile.NamedTemporaryFile() cmd3 = 'maq fastq2bfq %s %s 2>&1; maq fastq2bfq %s %s 2>&1; maq map -c %s.csmap %s %s %s 1>/dev/null 2>&1; maq mapview %s.csmap > %s.txt' %(fastq,tmpbfq_f.name,fastq_r,tmpbfq_r.name,fastq,ref_csbfa.name,tmpbfq_f.name,tmpbfq_r.name,fastq,fastq) subprocess_dict['sp'+str(ii+1)] = subprocess.Popen([cmd3],shell=True,stdout=subprocess.PIPE) ii += 1 while True: all_done = True for j,k in enumerate(subprocess_dict.keys()): if subprocess_dict['sp'+str(j+1)].wait() != 0: err = subprocess_dict['sp'+str(j+1)].communicate()[1] if err != None: stop_err("Mapping error: %s" %err) all_done = False if all_done: break cmdout = "for map in *.txt; do cat $map >> %s; done" %(out_fname) os.system(cmdout) tmpcsmap = tempfile.NamedTemporaryFile() cmd_cat_csmap = "for csmap in *.csmap; do cat $csmap >> %s; done" %(tmpcsmap.name) os.system(cmd_cat_csmap) tmppileup = tempfile.NamedTemporaryFile() cmdpileup = "maq pileup -m %s -q %s %s %s > %s" %(max_mismatch,min_mapqual,ref_bfa.name,tmpcsmap.name,tmppileup.name) os.system(cmdpileup) tmppileup.seek(0) print >> out_f2, "#chr\tposition\tref_nt\tcoverage\tSNP_count\tA_count\tT_count\tG_count\tC_count" for line in file(tmppileup.name): elems = line.strip().split() ref_nt = elems[2].capitalize() read_nt = elems[4] coverage = int(elems[3]) a,t,g,c = 0,0,0,0 ref_nt_count = 0 for ch in read_nt: ch = ch.capitalize() if ch not in ['A','T','G','C',',','.']: continue if ch in [',','.']: ch = ref_nt ref_nt_count += 1 try: nt_ind = ['A','T','G','C'].index(ch) if nt_ind == 0: a+=1 elif nt_ind == 1: t+=1 elif nt_ind == 2: g+=1 else: c+=1 except ValueError, we: print >>sys.stderr, we print >> out_f2, "%s\t%s\t%s\t%s\t%s\t%s" %("\t".join(elems[:4]),coverage-ref_nt_count,a,t,g,c) except Exception, er2: stop_err("Encountered error while mapping: %s" %(str(er2))) else: #single end reads tmpf = tempfile.NamedTemporaryFile() tmpfastq = tempfile.NamedTemporaryFile() cmd1 = "solid2fastq_modified.pl 'no' %s %s %s %s %s %s %s 2>&1" %(tmpf.name,None,None,f3_read_fname,f3_qual_fname,None,None) try: os.system(cmd1) os.system('gunzip -c %s >> %s' %(tmpf.name,tmpfastq.name)) tmpf.close() except: stop_err("Error converting data to fastq format.") #make a temp directory where the split fastq files will be stored try: split_dir = tempfile.mkdtemp() split_file_prefix = tempfile.mktemp(dir=split_dir) splitcmd = 'split -a 2 -l %d %s %s' %(32000000,tmpfastq.name,split_file_prefix) #32M lines correspond to 8M reads os.system(splitcmd) os.chdir(split_dir) for i,fastq in enumerate(os.listdir(split_dir)): tmpbfq = tempfile.NamedTemporaryFile() cmd3 = 'maq fastq2bfq %s %s 2>&1; maq map -c %s.csmap %s %s 1>/dev/null 2>&1; maq mapview %s.csmap > %s.txt' %(fastq,tmpbfq.name,fastq,ref_csbfa.name,tmpbfq.name,fastq,fastq) subprocess_dict['sp'+str(i+1)] = subprocess.Popen([cmd3],shell=True,stdout=subprocess.PIPE) while True: all_done = True for j,k in enumerate(subprocess_dict.keys()): if subprocess_dict['sp'+str(j+1)].wait() != 0: err = subprocess_dict['sp'+str(j+1)].communicate()[1] if err != None: stop_err("Mapping error: %s" %err) all_done = False if all_done: break cmdout = "for map in *.txt; do cat $map >> %s; done" %(out_fname) os.system(cmdout) tmpcsmap = tempfile.NamedTemporaryFile() cmd_cat_csmap = "for csmap in *.csmap; do cat $csmap >> %s; done" %(tmpcsmap.name) os.system(cmd_cat_csmap) tmppileup = tempfile.NamedTemporaryFile() cmdpileup = "maq pileup -m %s -q %s %s %s > %s" %(max_mismatch,min_mapqual,ref_bfa.name,tmpcsmap.name,tmppileup.name) os.system(cmdpileup) tmppileup.seek(0) print >> out_f2, "#chr\tposition\tref_nt\tcoverage\tSNP_count\tA_count\tT_count\tG_count\tC_count" for line in file(tmppileup.name): elems = line.strip().split() ref_nt = elems[2].capitalize() read_nt = elems[4] coverage = int(elems[3]) a,t,g,c = 0,0,0,0 ref_nt_count = 0 for ch in read_nt: ch = ch.capitalize() if ch not in ['A','T','G','C',',','.']: continue if ch in [',','.']: ch = ref_nt ref_nt_count += 1 try: nt_ind = ['A','T','G','C'].index(ch) if nt_ind == 0: a+=1 elif nt_ind == 1: t+=1 elif nt_ind == 2: g+=1 else: c+=1 except: pass print >> out_f2, "%s\t%s\t%s\t%s\t%s\t%s" %("\t".join(elems[:4]),coverage-ref_nt_count,a,t,g,c) except Exception, er2: stop_err("Encountered error while mapping: %s" %(str(er2))) #Build custom track from pileup chr_list=[] out_f2.seek(0) fcov = tempfile.NamedTemporaryFile() fout_a = tempfile.NamedTemporaryFile() fout_t = tempfile.NamedTemporaryFile() fout_g = tempfile.NamedTemporaryFile() fout_c = tempfile.NamedTemporaryFile() fcov.write('''track type=wiggle_0 name="Coverage track" description="Coverage track (from Galaxy)" color=0,0,0 visibility=2\n''') fout_a.write('''track type=wiggle_0 name="Track A" description="Track A (from Galaxy)" color=255,0,0 visibility=2\n''') fout_t.write('''track type=wiggle_0 name="Track T" description="Track T (from Galaxy)" color=0,255,0 visibility=2\n''') fout_g.write('''track type=wiggle_0 name="Track G" description="Track G (from Galaxy)" color=0,0,255 visibility=2\n''') fout_c.write('''track type=wiggle_0 name="Track C" description="Track C (from Galaxy)" color=255,0,255 visibility=2\n''') for line in out_f2: if line.startswith("#"): continue elems = line.split() chr = elems[0] if chr not in chr_list: chr_list.append(chr) if not (chr.startswith('chr') or chr.startswith('scaffold')): chr = 'chr' header = "variableStep chrom=%s" %(chr) fcov.write("%s\n" %(header)) fout_a.write("%s\n" %(header)) fout_t.write("%s\n" %(header)) fout_g.write("%s\n" %(header)) fout_c.write("%s\n" %(header)) try: pos = int(elems[1]) cov = int(elems[3]) a = int(elems[5]) t = int(elems[6]) g = int(elems[7]) c = int(elems[8]) except: continue fcov.write("%s\t%s\n" %(pos,cov)) try: a_freq = a*100./cov t_freq = t*100./cov g_freq = g*100./cov c_freq = c*100./cov except ZeroDivisionError: a_freq=t_freq=g_freq=c_freq=0 fout_a.write("%s\t%s\n" %(pos,a_freq)) fout_t.write("%s\t%s\n" %(pos,t_freq)) fout_g.write("%s\t%s\n" %(pos,g_freq)) fout_c.write("%s\t%s\n" %(pos,c_freq)) fcov.seek(0) fout_a.seek(0) fout_g.seek(0) fout_t.seek(0) fout_c.seek(0) os.system("cat %s %s %s %s %s | cat > %s" %(fcov.name,fout_a.name,fout_t.name,fout_g.name,fout_c.name,out_f3name)) if __name__=="__main__": __main__()