Rsamtools bam to ShortReadQ for export
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Marcus Davy ▴ 390
@marcus-davy-5153
Last seen 6.6 years ago
Hi, I would like to export certain reads filtered using Rsamtools, for example aligning against contaminant references to exclude a set of sequences filtering on the bitwise flag=0x4 (similar to SamToFastq.jar in the picard http://picard.sourceforge.net/ tools), but couldn't find much on exporting/writing to file in the Rsamtools-Overview.pdf vignette or on google. Is there a function/utility which converts an Rsamtools bam list region to ShortReadQ object(s) for export using writeFastq() with single end *and* paired end alignments which have been read in via the Rsamtools interface? ## Load a bam bam <- scanBam(...) ## Filter using R's subsetting, bamRegion <- bam[[1]][ ... ] ## list of list ## Should work for single end alignments, but not paired end alignments ## Read pairs may have to index on mpos = PNEXT field rfq <- ShortReadQ(bamRegion[["seq"]], bamRegion[["qual"]], BStringSet(bamRegion[["qname"]])) If not is the area of exporting from Rsamtools worth developing futher? cheers, Marcus [[alternative HTML version deleted]]
Rsamtools Rsamtools • 1.3k views
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@martin-morgan-1513
Last seen 3 months ago
United States
On 11/27/2012 01:17 PM, Marcus Davy wrote: > Hi, > I would like to export certain reads filtered using Rsamtools, for example > aligning against contaminant references > to exclude a set of sequences filtering on the bitwise flag=0x4 (similar to > SamToFastq.jar in the picard http://picard.sourceforge.net/ tools), > but couldn't find much on exporting/writing to file in the > Rsamtools-Overview.pdf vignette or on google. > > Is there a function/utility which converts an Rsamtools bam list region to > ShortReadQ object(s) for export using writeFastq() > with single end *and* paired end alignments which have been read in via the > Rsamtools interface? > > ## Load a bam > bam <- scanBam(...) > > ## Filter using R's subsetting, > bamRegion <- bam[[1]][ ... ] ## list of list > > ## Should work for single end alignments, but not paired end alignments > ## Read pairs may have to index on mpos = PNEXT field > rfq <- ShortReadQ(bamRegion[["seq"]], bamRegion[["qual"]], > BStringSet(bamRegion[["qname"]])) > > If not is the area of exporting from Rsamtools worth developing futher? I think I would use readBamGappedAlignments, specifying what=c("seq", "qual") and maybe "qname", flag=scanBamFlag(<...>) and perhaps the tag argument to filter / load relevant records, and then create a fastq object ShortReadQ(<...>) and writeFastq. This could be made space efficient using BamFile(, yieldSize=) and iterating through bf = open(BamFile(...)) out = FastqFile("..." while (nrow(ga <- readBamGappedAlignments(bf, ...))) { ## filter, create fq = ShortReadQ writeFastq(fq, out, mode ="a") } close(bf) close(out) > > cheers, > > Marcus > > [[alternative HTML version deleted]] > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor > -- Computational Biology / Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N. PO Box 19024 Seattle, WA 98109 Location: Arnold Building M1 B861 Phone: (206) 667-2793
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Thanks Martin, that gives me a new lead to explore further. Marcus On Wed, Nov 28, 2012 at 11:26 AM, Martin Morgan <mtmorgan@fhcrc.org> wrote: > On 11/27/2012 01:17 PM, Marcus Davy wrote: > >> Hi, >> I would like to export certain reads filtered using Rsamtools, for example >> aligning against contaminant references >> to exclude a set of sequences filtering on the bitwise flag=0x4 (similar >> to >> SamToFastq.jar in the picard http://picard.sourceforge.net/ tools), >> but couldn't find much on exporting/writing to file in the >> Rsamtools-Overview.pdf vignette or on google. >> >> Is there a function/utility which converts an Rsamtools bam list region to >> ShortReadQ object(s) for export using writeFastq() >> with single end *and* paired end alignments which have been read in via >> the >> Rsamtools interface? >> >> ## Load a bam >> bam <- scanBam(...) >> >> ## Filter using R's subsetting, >> bamRegion <- bam[[1]][ ... ] ## list of list >> >> ## Should work for single end alignments, but not paired end alignments >> ## Read pairs may have to index on mpos = PNEXT field >> rfq <- ShortReadQ(bamRegion[["seq"]], bamRegion[["qual"]], >> BStringSet(bamRegion[["qname"]**])) >> >> If not is the area of exporting from Rsamtools worth developing futher? >> > > I think I would use readBamGappedAlignments, specifying what=c("seq", > "qual") and maybe "qname", flag=scanBamFlag(<...>) and perhaps the tag > argument to filter / load relevant records, and then create a fastq object > ShortReadQ(<...>) and writeFastq. > > This could be made space efficient using BamFile(, yieldSize=) and > iterating through > > bf = open(BamFile(...)) > out = FastqFile("..." > while (nrow(ga <- readBamGappedAlignments(bf, ...))) { > ## filter, create fq = ShortReadQ > writeFastq(fq, out, mode ="a") > } > close(bf) > close(out) > > >> cheers, >> >> Marcus >> >> [[alternative HTML version deleted]] >> >> ______________________________**_________________ >> Bioconductor mailing list >> Bioconductor@r-project.org >> https://stat.ethz.ch/mailman/**listinfo/bioconductor<https: stat.e="" thz.ch="" mailman="" listinfo="" bioconductor=""> >> Search the archives: http://news.gmane.org/gmane.** >> science.biology.informatics.**conductor<http: news.gmane.org="" gmane="" .science.biology.informatics.conductor=""> >> >> > > -- > Computational Biology / Fred Hutchinson Cancer Research Center > 1100 Fairview Ave. N. > PO Box 19024 Seattle, WA 98109 > > Location: Arnold Building M1 B861 > Phone: (206) 667-2793 > [[alternative HTML version deleted]]
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