# HG changeset patch # User fubar # Date 1412038245 14400 # Node ID 624238a8440fea4900b24150b04dd5c94b52ca19 Uploaded diff -r 000000000000 -r 624238a8440f rg_rnaStar.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/rg_rnaStar.xml Mon Sep 29 20:50:45 2014 -0400 @@ -0,0 +1,289 @@ + + Gapped-read mapper for RNA-seq data + + rnastar + samtools + + + ## + ## Run STAR. + ## + + STAR + ## Can adjust this as appropriate for the system. + --genomeLoad NoSharedMemory + + --genomeDir ${refGenomeSource.index.fields.path} + --readFilesIn $input1 + #if $singlePaired.sPaired == "paired" + $singlePaired.input2 + #end if + --runThreadN 4 + #if $params.settingsType == "full": + --chimSegmentMin $params.chim_segment_min + --chimScoreMin $params.chim_score_min + #end if + + ## may or may not need to generate SAM tags and handle non-canonicals for Cufflinks tools. + ${outSAMstrandField} ${outFilterIntronMotifs} ${outSAMattributes} + + ; + + ## + ## BAM conversion. + ## + + ## Convert aligned reads. + samtools view -Shb Aligned.out.sam | samtools sort - Aligned.out + + ## Convert chimeric reads. + #if $params.settingsType == "full" and $params.chim_segment_min > 0: + ; samtools view -Shb Chimeric.out.sam | samtools sort - Chimeric.out + #end if + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + (params['settingsType'] == 'full' and params['chim_segment_min'] > 0) + + + + + + + + + + + + + + + + (params['settingsType'] == 'full' and params['chim_segment_min'] > 0) + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +**What it does** +Runs the rna star gapped aligner. Suited to paired or single end rna-seq. + +8.2: SAM alignments + +The number of loci Nmap a read maps to (multi-mapping) is given by NH:i: field. +The mapping quality MAPQ (column 5) is 255 for uniquely mapping reads, and int(-10*log10(1-1/Nmap)) for +multi-mapping reads. This scheme is same as the one used by Tophat and is compatible with Cufflinks. + +For multi-mappers, all alignments except one are marked with 0x100 (secondary alignment) in the FLAG +column 2. The un-marked alignment is either the best one (i.e. highest scoring), or is randomly selected from +the alignments of equal quality. + +8.2.1: Standard SAM attributes +With default --outSAMattributes Standard option the following SAM attributes will be generated: + +Column 12: NH: number of loci a read (pair) maps to +Column 13: IH: alignment index for all alignments of a read +Column 14: aS: alignment score +Column 15: nM: number of mismatches (does not include indels) + +8.2.2: Extra SAM attrbiutes +If --outSAMattributes All option is used, the following additional attributes will be output: + +Column 16: jM:B:c,M1,M2,... Intron motifs for all junctions (i.e. N in CIGAR): +0: non-canonical; 1:GT/AG, 2: CT/AC, 3: GC/AG, 4: CT/GC, 5: AT/AC, 6: GT/AT. + +If splice junctions database is used, and a junction is annotated, 20 is added to its motif value. +Column 17: jI:B:I,Start1,End1,Start2,End2,... Start and End of introns for all junctions (1-based) + +Note, that samtools 0.1.18 or later have to be used with these extra attributes. + + +8.2.3: XS SAM strand attribute for Cufflinks/Cuffdiff + +If you have un-stranded RNA-seq data, and wish to run Cufflinks/Cuffdiff on STAR alignments, you will +need to run STAR with --outSAMstrandField intronMotif option, which will generate the XS +strand attribute for all alignments that contain splice junctions. The spliced alignments that have undefined +strand (i.e. containing only non-canonical junctions) will be suppressed. + +If you have stranded RNA-seq data, you do not need to use any specific STAR options. Instead, you need +to run Cufflinks with the library option --library-type options. For example, cufflinks with +library-type fr-firststrand should be used for the “standard” dUTP protocol. +This option has to be used only for Cufflinks runs and not for STAR runs. + +It is recommended to remove the non-canonical junctions for Cufflinks runs using – + +--outFilterIntronMotifs RemoveNoncanonical +filter out alignments that contain non-canonical junctions + +OR + +--outFilterIntronMotifs RemoveNoncanonicalUnannotated +filter out alignments that contain non-canonical unannotated junctions +when using annotated splice junctions database. The annotated non- +canonical junctions will be kept. + + +**Attributions** + +Note that each component has its own license. Good luck with figuring out your obligations. + +rna_star - see the web site at rna_star_ + +For details, please see the rna_starMS_ +"STAR: ultrafast universal RNA-seq aligner" +A. Dobin et al, Bioinformatics 2012; doi: 10.1093/bioinformatics/bts635 + +Galaxy_ (that's what you are using right now!) for gluing everything together + +Most of the work for this wrapper XML is Jeremy Goecks' original STAR_ wrapper + +Minor tweaks to output names to suit our downstream purposes, toolshed automated dependencies +and odds and ends of other code and documentation comprising this tool was +written by Ross Lazarus and that part is licensed_ the same way as other rgenetics artefacts + +.. _STAR: https://bitbucket.org/jgoecks/jeremys-code/raw/fa1930a689b8e2f6b59cc1706e5ba0ed8ad357be/galaxy/tool-wrappers/star.xml +.. _licensed: http://creativecommons.org/licenses/by-nc-nd/3.0/ +.. _rna_star: http://code.google.com/p/rna-star/ +.. _rna_starMS: http://bioinformatics.oxfordjournals.org/content/29/1/15.full +.. _Galaxy: http://getgalaxy.org + + + diff -r 000000000000 -r 624238a8440f tool-data/rnastar_indexes.loc.sample --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tool-data/rnastar_indexes.loc.sample Mon Sep 29 20:50:45 2014 -0400 @@ -0,0 +1,11 @@ +#This is a sample file distributed with Galaxy that enables tools +#to use a directory of rna-star indexed sequences data files. You will +#need to create these data files and then create a bowtie_indices.loc +#file similar to this one (store it in this directory) that points to +#the directories in which those files are stored. The bowtie2_indices.loc +#file has this format (longer white space characters are TAB characters): +# +# +# +#hg19 hg19 hg19 full /mnt/galaxyIndices/genomes/hg19/rnastar + diff -r 000000000000 -r 624238a8440f tool_data_table_conf.xml.sample --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tool_data_table_conf.xml.sample Mon Sep 29 20:50:45 2014 -0400 @@ -0,0 +1,7 @@ + + + + value, dbkey, name, path + +
+
diff -r 000000000000 -r 624238a8440f tool_dependencies.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/tool_dependencies.xml Mon Sep 29 20:50:45 2014 -0400 @@ -0,0 +1,15 @@ + + + + + + + + + + +Installs the STAR wrapper and dependency packages samtools and star - see https://code.google.com/p/rna-star/ +STAR is a very fast mapper for rna-seq giving junctions if the indexes are constructed with a junction library + + +