details: http://www.bx.psu.edu/hg/galaxy/rev/25d736f5cbf7 changeset: 2409:25d736f5cbf7 user: guru date: Wed May 13 21:54:45 2009 -0400 description: MAQ mapping pipeline for SOLiD data 4 file(s) affected in this change: tool_conf.xml.sample tools/solid_tools/maq_cs_wrapper.py tools/solid_tools/maq_cs_wrapper.xml tools/solid_tools/maq_cs_wrapper_code.py diffs (418 lines): diff -r 3af0dea9a5c7 -r 25d736f5cbf7 tool_conf.xml.sample --- a/tool_conf.xml.sample Mon May 11 17:02:54 2009 -0400 +++ b/tool_conf.xml.sample Wed May 13 21:54:45 2009 -0400 @@ -284,6 +284,7 @@ <section name="SOLiD Data Analysis" id="solid_tools"> <tool file="solid_tools/solid_qual_stats.xml" /> <tool file="solid_tools/solid_qual_boxplot.xml" /> + <tool file="solid_tools/maq_cs_wrapper.xml" /> </section> <section name="FASTA manipulation" id="fasta_manipulation"> <tool file="fasta_tools/fasta_compute_length.xml" /> diff -r 3af0dea9a5c7 -r 25d736f5cbf7 tools/solid_tools/maq_cs_wrapper.py --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tools/solid_tools/maq_cs_wrapper.py Wed May 13 21:54:45 2009 -0400 @@ -0,0 +1,270 @@ +#! /usr/bin/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('zcat -f %s >> %s' %(tmpf.name,tmpffastq.name)) + os.system('zcat -f %s >> %s' %(tmpr.name,tmprfastq.name)) + os.system('zcat -f %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('zcat -f %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__() + + diff -r 3af0dea9a5c7 -r 25d736f5cbf7 tools/solid_tools/maq_cs_wrapper.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tools/solid_tools/maq_cs_wrapper.xml Wed May 13 21:54:45 2009 -0400 @@ -0,0 +1,120 @@ +<tool id="maq_cs_wrapper" name="MAQ for SOLiD" version="1.0.0"> + <description> </description> + <command interpreter="python"> + maq_cs_wrapper.py + $output1 + $output2 + $ref + $library_type.f3_reads + $library_type.f3_qual + $library_type.is_paired + #if $library_type.is_paired == "yes": + $library_type.r3_reads + $library_type.r3_qual + #else: + "None" + "None" + #end if + $min_mapqual + $max_mismatch + $output3 + + </command> + + <inputs> + <param name="ref" type="data" format="fasta" label="Target Genome"/> + <conditional name="library_type"> + <param name="is_paired" type="select" label="Is the library mate-paired?" multiple="false"> + <option value="no">No</option> + <option value="yes">Yes</option> + </param> + <when value="no"> + <param name="f3_reads" type="data" format="csfasta" label="F3 reads file"/> + <param format="qualsolid" name="f3_qual" type="data" label="F3 quality file" help="If your dataset doesn't show up in the menu, click the pencil icon next to your dataset and set the datatype to 'qualsolid'" /> + </when> + <when value="yes"> + <param name="f3_reads" type="data" format="csfasta" label="F3 reads file"/> + <param format="qualsolid" name="f3_qual" type="data" label="F3 quality file" help="If your dataset doesn't show up in the menu, click the pencil icon next to your dataset and set the datatype to 'qualsolid'" /> + <param name="r3_reads" type="data" format="csfasta" label="R3 reads file"/> + <param format="qualsolid" name="r3_qual" type="data" label="R3 quality file" help="If your dataset doesn't show up in the menu, click the pencil icon next to your dataset and set the datatype to 'qualsolid'" /> + </when> + </conditional> + <param name="min_mapqual" type="integer" size="3" value="0" label="Minimum mapping quality allowed for a read to be used" help="Reads below the specified mapping quality will not be considered in coverage and SNP analysis."/> + <param name="max_mismatch" type="integer" size="3" value="7" label="Maximum number of mismatches allowed for a read to be used" help="Reads above the specified threshold will not be considered in coverage and SNP analysis."/> + </inputs> + <outputs> + <data format="tabular" name="output1" metadata_source="ref" /> + <data format="tabular" name="output2" metadata_source="ref" /> + <data format="customtrack" name="output3" metadata_source="ref" /> + </outputs> + + <!-- "ToolTestCase does not deal with multiple outputs properly yet." + <tests> + + <test> + <param name="ref" value="phiX_mod.fasta" /> + <param name="is_paired" value="no" /> + <param name="f3_reads" value="phiX_solid.csfasta" /> + <param name="f3_qual" value="phiX_solid.qualsolid" /> + <param name="min_mapqual" value="0" /> + <param name="max_mismatch" value="7" /> + <output name="output1" file="phiX_solid_maq.map" /> + <output name="output2" file="phiX_solid_maq.pileup" /> + <output name="output3" file="phiX_solid_maq.ctrack" /> + + </test> + </tests> + --> +<help> + +.. class:: infomark + +**What it does** + +This tool maps SOLiD colour-space reads against the target genome using MAQ. It produces three output datasets: + + +**ALIGNMENT INFO** : contains the read alignment information, + +**PILEUP** : contains the coverage and SNP statistics for every nucleotide of the target genome, + +**CUSTOM TRACK** : contains the coverage and SNP statistics as custom tracks displayable in the UCSC browser. + +----- + +**The ALIGNMENT INFO dataset will contain the following fields:** + +* column 1 = read name +* column 2 = chromosome +* column 3 = position +* column 4 = strand +* column 5 = insert size from the outer coorniates of a pair +* column 6 = paired flag +* column 7 = mapping quality +* column 8 = single-end mapping quality +* column 9 = alternative mapping quality +* column 10 = number of mismatches of the best hit +* column 11 = sum of qualities of mismatched bases of the best hit +* column 12 = number of 0-mismatch hits of the first 24bp +* column 13 = number of 1-mismatch hits of the first 24bp on the reference +* column 14 = length of the read +* column 15 = read sequence +* column 16 = read quality + + +**The PILEUP dataset will contain the following fields:** + +* column 1 = chromosome +* column 2 = position +* column 3 = reference nucleotide +* column 4 = coverage (number of reads that cover this position) +* column 5 = number of SNPs +* column 6 = number of As +* column 7 = number of Ts +* column 8 = number of Gs +* column 9 = number of Cs + +</help> +<code file="maq_cs_wrapper_code.py"/> + +</tool> diff -r 3af0dea9a5c7 -r 25d736f5cbf7 tools/solid_tools/maq_cs_wrapper_code.py --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tools/solid_tools/maq_cs_wrapper_code.py Wed May 13 21:54:45 2009 -0400 @@ -0,0 +1,5 @@ +def exec_before_job(app, inp_data, out_data, param_dict, tool): + out_data['output1'].name = out_data['output1'].name + " [ ALIGNMENT INFO ]" + out_data['output2'].name = out_data['output2'].name + " [ PILEUP ]" + out_data['output3'].name = out_data['output3'].name + " [ CUSTOM TRACK ]" +