Hi,
I've a little problem to understand the scripture command line :
So I've the accepted_hits.bam for all my samples. So I have to merge all the bam files into one big bam file before running scripture. In the help file there is two alignment file param :
-alignment --pairedEnd
Which one do I have to use to input the merged bam file ?
Here's the help file :
Parameters
-alignment <Alignment file in BAM, SAM or Alignemnt format>
-maskFileDir <Mask File directory>
-out <Output file name>
-windows <Comma separated list of windows to evaluate>
-sizeFile <Chromosome size file>
Optional arguments:
-trim <Include this flag if trimming of the ends of windows based on read coverage is desired this is expensive>
-alpha <Desired FDR>
one of -chr <Chromsomosome to segment> or -chrSequence <Necessary to filter spliced reads by splice site information. Notice that this is only compatible with region files that contain regions of only one chromosome> -dontFilterCanonicalSplice
-start <To segment only a subregion of the chromosome include its start> -end <To segment only a subregion of the chromosome include its end>
-minSpliceSupport <Minimum count to support splice reads, default is 1>
-pairedEnd <Paired end alignment files> -strandSpecificReads <Strand specific alignment file> -scoreRegions <Full BED to score> -upWeightSplices -lambda <If a prior background expectation for number of reads per base exists> -exons <BED file of exons> -introns <Introns and counts>
Task: AddPairs - Uses a paired end alignment to tune graph
-in <Graph in .dot format. Standard input is assumed>
-pairedEnd <Paired end information (as in previous task), in single line BED format>
-maskFileDir <Directory containing mask files for the genome>
-chr <Chromosome (only a chromosome at a time is supported at this point)>
-sizeFile <Chromosome size file>
-out <Output file name>
Task: score - Computes several expression related scores for a set of annotations -in <Full BED file with annotations to score>
-alignment <Alignment file in BAM, SAM or Alignemnt format>
-sizeFile <Chromosome size file>
-out <Output file name>
-maskFileDir <Mask File directory>
Thanks,
N.
1 answer
I just read about this tool and will share what I understood. So you can read a walkthrough example here.So if you have paired end data, then you have option to use pairedEnd argument. This argument will take files generated by makePairedFile task.
From the link- So if you have paired end data, you will have 2 files for each "accepted_hits.sorted.sam". First you have to use makePairedFile task-
>> java -Xmx4000m -jar scripture.jar -task makePairedFile -pair1 tophat_out_SRR039999_1/accepted_hits.sorted.sam -pair2 tophat_out_SRR039999_2/accepted_hits.sorted.sam -out SRR039999.paired.sam -sorted
>> java -Xmx4000m -jar scripture.jar -task makePairedFile -pair1 tophat_out_SRR040000_1/accepted_hits.sorted.sam -pair2 tophat_out_SRR040000_2/accepted_hits.sorted.sam -out SRR040000.paired.sam -sorted
>> java -Xmx4000m -jar scripture.jar -task makePairedFile -pair1 tophat_out_SRR040001_1/accepted_hits.sorted.sam -pair2 tophat_out_SRR040001_2/accepted_hits.sorted.sam -out SRR040001.paired.sam -sorted
and then combine these files-
>> cat SRR039999.paired.sam SRR040000.paired.sam SRR040001.paired.sam > all_alignments.paired.sam
and then sort and index. So now this file "all_alignments.paired.sorted.sam" will be used in pairedEnd argument. and for -alignment parameter you will simple combine your bam files-
>> cat tophat_out_SRR039999_1/accepted_hits.sorted.sam tophat_out_SRR039999_2/accepted_hits.sorted.sam tophat_out_SRR040000_1/accepted_hits.sorted.sam tophat_out_SRR040000_2/accepted_hits.sorted.sam tophat_out_SRR040001_1/accepted_hits.sorted.sam tophat_out_SRR040001_2/accepted_hits.sorted.sam > all_alignments.sam
Sort and index.Now this "all_alignments.sorted.sam" will be used in -alignment parameter.
So they used this command in example-
>>java –jar scripture.jar –alignment all_alignments.sorted.sam –out chr19.scriptureESTest.segments –sizeFile mm9.sizes –chr chr19 –chrSequence chr19.fa -pairedEnd all_alignments.paired.sorted.sam
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