From 8986dc2c812a22489c930f55366a9bbcb08ebab1 Mon Sep 17 00:00:00 2001 From: Daniel Blankenberg Date: Mon, 21 Nov 2011 16:30:34 -0500 Subject: [PATCH] Add SAMTools mpileup. --- datatypes_conf.xml.sample | 1 + tool_conf.xml.sample | 1 + tool_data_table_conf.xml.sample | 5 + tools/samtools/samtools_mpileup.xml | 213 ++++++++++++++++++++++++++++ tools/samtools/samtools_wrapper.py | 110 ++++++++++++++ 5 files changed, 330 insertions(+) create mode 100644 tools/samtools/samtools_mpileup.xml create mode 100644 tools/samtools/samtools_wrapper.py diff --git a/datatypes_conf.xml.sample b/datatypes_conf.xml.sample index cd87e71aae0..5f756eec957 100644 --- a/datatypes_conf.xml.sample +++ b/datatypes_conf.xml.sample @@ -159,6 +159,7 @@ + diff --git a/tool_conf.xml.sample b/tool_conf.xml.sample index ce4347e756f..ee3ba7e8ba1 100644 --- a/tool_conf.xml.sample +++ b/tool_conf.xml.sample @@ -371,6 +371,7 @@ + diff --git a/tool_data_table_conf.xml.sample b/tool_data_table_conf.xml.sample index a217776b72a..5a899e0990b 100644 --- a/tool_data_table_conf.xml.sample +++ b/tool_data_table_conf.xml.sample @@ -65,6 +65,11 @@ value, name, path + + + line_type, value, path + +
value, dbkey, name, path diff --git a/tools/samtools/samtools_mpileup.xml b/tools/samtools/samtools_mpileup.xml new file mode 100644 index 00000000000..6068e1efaea --- /dev/null +++ b/tools/samtools/samtools_mpileup.xml @@ -0,0 +1,213 @@ + + SNP and indel caller + + samtools + + samtools_wrapper.py + -p 'samtools mpileup' + --stdout "${output_log}" + #if $reference_source.reference_source_selector != "history": + -p '-f "${reference_source.ref_file.fields.path}"' + #else: + -d "-f" "${reference_source.ref_file}" "fa" "reference_input" + #end if + #for $i, $input_bam in enumerate( $reference_source.input_bams ): + -d " " "${input_bam.input_bam}" "${input_bam.input_bam.ext}" "bam_input_${i}" + -d "" "${input_bam.input_bam.metadata.bam_index}" "bam_index" "bam_input_${i}" ##hardcode galaxy ext type as bam_index + #end for + -p ' + #if str( $advanced_options.advanced_options_selector ) == "advanced": + ${advanced_options.skip_anomalous_read_pairs} + ${advanced_options.disable_probabilistic_realignment} + -C "${advanced_options.coefficient_for_downgrading}" + -d "${advanced_options.max_reads_per_bam}" + ${advanced_options.extended_BAQ_computation} + #if str( $advanced_options.position_list ) != 'None': + -l "${advanced_options.position_list}" + #end if + -q "${advanced_options.minimum_mapping_quality}" + -Q "${advanced_options.minimum_base_quality}" + #if str( $advanced_options.region_string ): + -r "${advanced_options.region_string}" + #end if + ${advanced_options.output_per_sample_read_depth} + ${advanced_options.output_per_sample_strand_bias_p_value} + #end if + #if str( $genotype_likelihood_computation_type.genotype_likelihood_computation_type_selector ) == 'perform_genotype_likelihood_computation': + ##-g or -u + -g + -e "${genotype_likelihood_computation_type.gap_extension_sequencing_error_probability}" + -h "${genotype_likelihood_computation_type.coefficient_for_modeling_homopolymer_errors}" + #if str( $genotype_likelihood_computation_type.perform_indel_calling.perform_indel_calling_selector ) == 'perform_indel_calling': + -L "${genotype_likelihood_computation_type.perform_indel_calling.skip_indel_calling_above_sample_depth}" + #else: + -I + #end if + -o "${genotype_likelihood_computation_type.gap_open_sequencing_error_probability}" + #if len( $genotype_likelihood_computation_type.platform_list_repeat ): + -P "${ ",".join( [ str( platform.platform_entry ) for platform in $genotype_likelihood_computation_type.platform_list_repeat ] ) }" + #end if + #end if + > "${output_mpileup}" + ' + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +**What it does** + + Generate BCF or pileup for one or multiple BAM files. Alignment records are grouped by sample identifiers in @RG header lines. If sample identifiers are absent, each input file is regarded as one sample. + +------ + +**Settings**:: + + Input Options: + -6 Assume the quality is in the Illumina 1.3+ encoding. + -A Do not skip anomalous read pairs in variant calling. + -B Disable probabilistic realignment for the computation of base alignment quality (BAQ). BAQ is the Phred-scaled probability of a read base being misaligned. Applying this option greatly helps to reduce false SNPs caused by misalignments. + -b FILE List of input BAM files, one file per line [null] + -C INT Coefficient for downgrading mapping quality for reads containing excessive mismatches. Given a read with a phred-scaled probability q of being generated from the mapped position, the new mapping quality is about sqrt((INT-q)/INT)*INT. A zero value disables this functionality; if enabled, the recommended value for BWA is 50. [0] + -d INT At a position, read maximally INT reads per input BAM. [250] + -E Extended BAQ computation. This option helps sensitivity especially for MNPs, but may hurt specificity a little bit. + -f FILE The faidx-indexed reference file in the FASTA format. The file can be optionally compressed by razip. [null] + -l FILE BED or position list file containing a list of regions or sites where pileup or BCF should be generated [null] + -q INT Minimum mapping quality for an alignment to be used [0] + -Q INT Minimum base quality for a base to be considered [13] + -r STR Only generate pileup in region STR [all sites] + Output Options: + + -D Output per-sample read depth + -g Compute genotype likelihoods and output them in the binary call format (BCF). + -S Output per-sample Phred-scaled strand bias P-value + -u Similar to -g except that the output is uncompressed BCF, which is preferred for piping. + + Options for Genotype Likelihood Computation (for -g or -u): + + -e INT Phred-scaled gap extension sequencing error probability. Reducing INT leads to longer indels. [20] + -h INT Coefficient for modeling homopolymer errors. Given an l-long homopolymer run, the sequencing error of an indel of size s is modeled as INT*s/l. [100] + -I Do not perform INDEL calling + -L INT Skip INDEL calling if the average per-sample depth is above INT. [250] + -o INT Phred-scaled gap open sequencing error probability. Reducing INT leads to more indel calls. [40] + -P STR Comma dilimited list of platforms (determined by @RG-PL) from which indel candidates are obtained. It is recommended to collect indel candidates from sequencing technologies that have low indel error rate such as ILLUMINA. [all] + +------ + +**Citation** + +For the underlying tool, please cite `Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R; 1000 Genome Project Data Processing Subgroup. The Sequence Alignment/Map format and SAMtools. Bioinformatics. 2009 Aug 15;25(16):2078-9. <http://www.ncbi.nlm.nih.gov/pubmed/19505943>`_ + +If you use this tool in Galaxy, please cite Blankenberg D, et al. *In preparation.* + + + diff --git a/tools/samtools/samtools_wrapper.py b/tools/samtools/samtools_wrapper.py new file mode 100644 index 00000000000..ade066b0b7b --- /dev/null +++ b/tools/samtools/samtools_wrapper.py @@ -0,0 +1,110 @@ +#!/usr/bin/env python +#Dan Blankenberg + +""" +A wrapper script for running SAMTools commands. +""" + +import sys, optparse, os, tempfile, subprocess, shutil +from string import Template + +GALAXY_EXT_TO_SAMTOOLS_EXT = { 'bam_index':'bam.bai', } #items not listed here will use the galaxy extension as-is +GALAXY_EXT_TO_SAMTOOLS_FILE_TYPE = GALAXY_EXT_TO_SAMTOOLS_EXT #for now, these are the same, but could be different if needed +DEFAULT_SAMTOOLS_PREFIX = "SAMTools_file" +CHUNK_SIZE = 2**20 #1mb + + +def cleanup_before_exit( tmp_dir ): + if tmp_dir and os.path.exists( tmp_dir ): + shutil.rmtree( tmp_dir ) + +def SAMTOOLS_filename_from_galaxy( galaxy_filename, galaxy_ext, target_dir = None, prefix = None ): + suffix = GALAXY_EXT_TO_SAMTOOLS_EXT.get( galaxy_ext, galaxy_ext ) + if prefix is None: + prefix = DEFAULT_SAMTOOLS_PREFIX + if target_dir is None: + target_dir = os.getcwd() + SAMTools_filename = os.path.join( target_dir, "%s.%s" % ( prefix, suffix ) ) + os.symlink( galaxy_filename, SAMTools_filename ) + return SAMTools_filename + +def SAMTOOLS_filetype_argument_substitution( argument, galaxy_ext ): + return argument % dict( file_type = GALAXY_EXT_TO_SAMTOOLS_FILE_TYPE.get( galaxy_ext, galaxy_ext ) ) + +def open_file_from_option( filename, mode = 'rb' ): + if filename: + return open( filename, mode = mode ) + return None + +def html_report_from_directory( html_out, dir ): + html_out.write( '\n\nGalaxy - SAMTOOLS Output\n\n\n

\n

    \n' ) + for fname in sorted( os.listdir( dir ) ): + html_out.write( '
  • %s
    • \n' % ( fname, fname ) ) + html_out.write( '
\n\n\n' ) + +def __main__(): + #Parse Command Line + parser = optparse.OptionParser() + parser.add_option( '-p', '--pass_through', dest='pass_through_options', action='append', type="string", help='These options are passed through directly to SAMTOOLS, without any modification.' ) + parser.add_option( '-d', '--dataset', dest='datasets', action='append', type="string", nargs=4, help='"-argument" "original_filename" "galaxy_filetype" "name_prefix"' ) + parser.add_option( '', '--stdout', dest='stdout', action='store', type="string", default=None, help='If specified, the output of stdout will be written to this file.' ) + parser.add_option( '', '--stderr', dest='stderr', action='store', type="string", default=None, help='If specified, the output of stderr will be written to this file.' ) + parser.add_option( '', '--html_report_from_directory', dest='html_report_from_directory', action='append', type="string", nargs=2, help='"Target HTML File" "Directory"') + (options, args) = parser.parse_args() + + tmp_dir = tempfile.mkdtemp( prefix='tmp-SAMTOOLS-' ) + + #set up stdout and stderr output options + stdout = open_file_from_option( options.stdout, mode = 'wb' ) + stderr = open_file_from_option( options.stderr, mode = 'wb' ) + #if no stderr file is specified, we'll use our own + if stderr is None: + stderr = tempfile.NamedTemporaryFile( prefix="SAMTOOLS-stderr-", dir=tmp_dir ) + + if options.pass_through_options: + cmd = ' '.join( options.pass_through_options ) + else: + cmd = '' + return_code = None + if options.datasets: + for ( dataset_arg, filename, galaxy_ext, prefix ) in options.datasets: + SAMTools_filename = SAMTOOLS_filename_from_galaxy( filename, galaxy_ext, target_dir = tmp_dir, prefix = prefix ) + if dataset_arg: + if '>' in cmd: + cmd = cmd.replace( '>', ' %s "%s" >' % ( SAMTOOLS_filetype_argument_substitution( dataset_arg, galaxy_ext ), SAMTools_filename ), 1 ) + else: + cmd = '%s %s "%s"' % ( cmd, SAMTOOLS_filetype_argument_substitution( dataset_arg, galaxy_ext ), SAMTools_filename ) + #auto index fasta files: + if galaxy_ext == 'fa': + index_cmd = 'samtools faidx %s' % ( SAMTools_filename ) + proc = subprocess.Popen( args=index_cmd, stdout=stdout, stderr=stderr, shell=True, cwd=tmp_dir ) + return_code = proc.wait() + if return_code: + break + if return_code is None or not return_code: + proc = subprocess.Popen( args=cmd, stdout=stdout, stderr=stderr, shell=True, cwd=tmp_dir ) + return_code = proc.wait() + if return_code: + stderr_target = sys.stderr + else: + if stdout: + stderr_target = stdout + else: + stderr_target = sys.stdout + stderr.flush() + stderr.seek(0) + while True: + chunk = stderr.read( CHUNK_SIZE ) + if chunk: + stderr_target.write( chunk ) + else: + break + stderr.close() + #generate html reports + if options.html_report_from_directory: + for ( html_filename, html_dir ) in options.html_report_from_directory: + html_report_from_directory( open( html_filename, 'wb' ), html_dir ) + + cleanup_before_exit( tmp_dir ) + +if __name__=="__main__": __main__()