pileupAsGRanges depth limit
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Mark Dunning ★ 1.1k
@mark-dunning-3319
Last seen 13 months ago
Sheffield, Uk
Hi all, I'm running the pileupAsGRanges function from ggbio, and just wondering if there was a limit to the depth it reports? I have targeted sequencing data and expect many thousands of reads per base but the depth seems to be around 250? > pgr[3:5, + ] GRanges with 3 ranges and 7 metadata columns: seqnames ranges strand | A C G <rle> <iranges> <rle> | <integer> <integer> <integer> [1] chr17 [7577489, 7577489] + | 1 0 249 [2] chr17 [7577490, 7577490] + | 0 6 245 [3] chr17 [7577491, 7577491] + | 1 250 0 T N depth bam <integer> <integer> <numeric> <character> [1] 0 0 250 CleanedBams/FLD0097.bam [2] 0 0 251 CleanedBams/FLD0097.bam [3] 1 0 252 CleanedBams/FLD0097.bam I think that samtools mpileup has a default depth of 250, so is it the case that the applyPileup function is using the same limit, and it is possible to change it? Mark
Sequencing ggbio Sequencing ggbio • 1.4k views
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@martin-morgan-1513
Last seen 3 days ago
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On 12/17/2012 03:12 AM, Mark Dunning wrote: > Hi all, > > I'm running the pileupAsGRanges function from ggbio, and just > wondering if there was a limit to the depth it reports? I have > targeted sequencing data and expect many thousands of reads per base > but the depth seems to be around 250? Hi Mark -- applyPileups takes a 'PileupParam' argument, which has maxDepth=250L as an argument, so maxDepth = .Machine$integer.max would get you everything. The ... arguments of pileupAsGRanges gets passed to PileupParam, so I think you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). Martin > >> pgr[3:5, > + ] > GRanges with 3 ranges and 7 metadata columns: > seqnames ranges strand | A C G > <rle> <iranges> <rle> | <integer> <integer> <integer> > [1] chr17 [7577489, 7577489] + | 1 0 249 > [2] chr17 [7577490, 7577490] + | 0 6 245 > [3] chr17 [7577491, 7577491] + | 1 250 0 > T N depth bam > <integer> <integer> <numeric> <character> > [1] 0 0 250 CleanedBams/FLD0097.bam > [2] 0 0 251 CleanedBams/FLD0097.bam > [3] 1 0 252 CleanedBams/FLD0097.bam > > > I think that samtools mpileup has a default depth of 250, so is it the > case that the applyPileup function is using the same limit, and it is > possible to change it? > > Mark > > _______________________________________________ > 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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Do you guys think that pileupAsGRanges should be pushed up somewhere, outside of ggbio, or is it too high-level? There are clearly many ways that the pileup output could be coerced to GRanges; maybe Rsamtools or GenomicRanges could try to support some of the most useful? Michael On Mon, Dec 17, 2012 at 9:22 AM, Martin Morgan <mtmorgan@fhcrc.org> wrote: > On 12/17/2012 03:12 AM, Mark Dunning wrote: > >> Hi all, >> >> I'm running the pileupAsGRanges function from ggbio, and just >> wondering if there was a limit to the depth it reports? I have >> targeted sequencing data and expect many thousands of reads per base >> but the depth seems to be around 250? >> > > Hi Mark -- > > applyPileups takes a 'PileupParam' argument, which has maxDepth=250L as an > argument, so maxDepth = .Machine$integer.max would get you everything. > > The ... arguments of pileupAsGRanges gets passed to PileupParam, so I > think you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). > > Martin > > > >> pgr[3:5, >>> >> + ] >> GRanges with 3 ranges and 7 metadata columns: >> seqnames ranges strand | A C G >> <rle> <iranges> <rle> | <integer> <integer> <integer> >> [1] chr17 [7577489, 7577489] + | 1 0 249 >> [2] chr17 [7577490, 7577490] + | 0 6 245 >> [3] chr17 [7577491, 7577491] + | 1 250 0 >> T N depth bam >> <integer> <integer> <numeric> <character> >> [1] 0 0 250 CleanedBams/FLD0097.bam >> [2] 0 0 251 CleanedBams/FLD0097.bam >> [3] 1 0 252 CleanedBams/FLD0097.bam >> >> >> I think that samtools mpileup has a default depth of 250, so is it the >> case that the applyPileup function is using the same limit, and it is >> possible to change it? >> >> Mark >> >> ______________________________**_________________ >> 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 > > > ______________________________**_________________ > Bioconductor mailing list > Bioconductor@r-project.org > https://stat.ethz.ch/mailman/**listinfo/bioconductor<https: stat.et="" hz.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=""> > [[alternative HTML version deleted]]
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Hi Michael, Just a friendly reminder, pileupAsGRanges is not implemented in ggbio, it's currently in biovizBase, but I agree with you that this function might be more appropriate in some other packages like Rsamtools? Tengfei On Mon, Dec 17, 2012 at 12:13 PM, Michael Lawrence < lawrence.michael@gene.com> wrote: > Do you guys think that pileupAsGRanges should be pushed up somewhere, > outside of ggbio, or is it too high-level? There are clearly many ways that > the pileup output could be coerced to GRanges; maybe Rsamtools or > GenomicRanges could try to support some of the most useful? > > Michael > > > > > On Mon, Dec 17, 2012 at 9:22 AM, Martin Morgan <mtmorgan@fhcrc.org> wrote: > > > On 12/17/2012 03:12 AM, Mark Dunning wrote: > > > >> Hi all, > >> > >> I'm running the pileupAsGRanges function from ggbio, and just > >> wondering if there was a limit to the depth it reports? I have > >> targeted sequencing data and expect many thousands of reads per base > >> but the depth seems to be around 250? > >> > > > > Hi Mark -- > > > > applyPileups takes a 'PileupParam' argument, which has maxDepth=250L as > an > > argument, so maxDepth = .Machine$integer.max would get you everything. > > > > The ... arguments of pileupAsGRanges gets passed to PileupParam, so I > > think you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). > > > > Martin > > > > > > > >> pgr[3:5, > >>> > >> + ] > >> GRanges with 3 ranges and 7 metadata columns: > >> seqnames ranges strand | A C > G > >> <rle> <iranges> <rle> | <integer> <integer> > <integer> > >> [1] chr17 [7577489, 7577489] + | 1 0 > 249 > >> [2] chr17 [7577490, 7577490] + | 0 6 > 245 > >> [3] chr17 [7577491, 7577491] + | 1 250 > 0 > >> T N depth bam > >> <integer> <integer> <numeric> <character> > >> [1] 0 0 250 CleanedBams/FLD0097.bam > >> [2] 0 0 251 CleanedBams/FLD0097.bam > >> [3] 1 0 252 CleanedBams/FLD0097.bam > >> > >> > >> I think that samtools mpileup has a default depth of 250, so is it the > >> case that the applyPileup function is using the same limit, and it is > >> possible to change it? > >> > >> Mark > >> > >> ______________________________**_________________ > >> Bioconductor mailing list > >> Bioconductor@r-project.org > >> https://stat.ethz.ch/mailman/**listinfo/bioconductor< > https://stat.ethz.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 > > > > > > ______________________________**_________________ > > Bioconductor mailing list > > Bioconductor@r-project.org > > https://stat.ethz.ch/mailman/**listinfo/bioconductor< > https://stat.ethz.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> > > > > [[alternative HTML version deleted]] > > _______________________________________________ > Bioconductor mailing list > Bioconductor@r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: > http://news.gmane.org/gmane.science.biology.informatics.conductor > -- Tengfei Yin MCDB PhD student 1620 Howe Hall, 2274, Iowa State University Ames, IA,50011-2274 [[alternative HTML version deleted]]
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On 12/17/2012 10:13 AM, Michael Lawrence wrote: > Do you guys think that pileupAsGRanges should be pushed up somewhere, outside of > ggbio, or is it too high-level? There are clearly many ways that the pileup > output could be coerced to GRanges; maybe Rsamtools or GenomicRanges could try > to support some of the most useful? I'm ok with moving this (or some similar functionality) to Rsamtools; is the output of pileupAsGRanges what you're interested in, or could it be more useful? > > Michael > > > > > On Mon, Dec 17, 2012 at 9:22 AM, Martin Morgan <mtmorgan at="" fhcrc.org=""> <mailto:mtmorgan at="" fhcrc.org="">> wrote: > > On 12/17/2012 03:12 AM, Mark Dunning wrote: > > Hi all, > > I'm running the pileupAsGRanges function from ggbio, and just > wondering if there was a limit to the depth it reports? I have > targeted sequencing data and expect many thousands of reads per base > but the depth seems to be around 250? > > > Hi Mark -- > > applyPileups takes a 'PileupParam' argument, which has maxDepth=250L as an > argument, so maxDepth = .Machine$integer.max would get you everything. > > The ... arguments of pileupAsGRanges gets passed to PileupParam, so I think > you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). > > Martin > > > > pgr[3:5, > > + ] > GRanges with 3 ranges and 7 metadata columns: > seqnames ranges strand | A C G > <rle> <iranges> <rle> | <integer> <integer> <integer> > [1] chr17 [7577489, 7577489] + | 1 0 249 > [2] chr17 [7577490, 7577490] + | 0 6 245 > [3] chr17 [7577491, 7577491] + | 1 250 0 > T N depth bam > <integer> <integer> <numeric> <character> > [1] 0 0 250 CleanedBams/FLD0097.bam > [2] 0 0 251 CleanedBams/FLD0097.bam > [3] 1 0 252 CleanedBams/FLD0097.bam > > > I think that samtools mpileup has a default depth of 250, so is it the > case that the applyPileup function is using the same limit, and it is > possible to change it? > > Mark > > _________________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org <mailto:bioconductor at="" r-project.org=""> > https://stat.ethz.ch/mailman/__listinfo/bioconductor > <https: stat.ethz.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 <tel:%28206%29%20667-2793> > > > _________________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org <mailto:bioconductor at="" r-project.org=""> > https://stat.ethz.ch/mailman/__listinfo/bioconductor > <https: stat.ethz.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
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Hmm. It might be best to wait. We've got this other bam_tally pileup generator in gmapR. It has some features that applyPileups lacks. These would include: 1) per-cycle (read position) information 2) per-strand information 3) indels Note that indels are only available from the latest devel with indels=TRUE. Still very much in flux. I like the callback-based applyPileups API. So there are good things about each, as well as a lot of overlap. How hard would it be to add those features to applyPileups? Michael On Mon, Dec 17, 2012 at 1:19 PM, Martin Morgan <mtmorgan@fhcrc.org> wrote: > On 12/17/2012 10:13 AM, Michael Lawrence wrote: > >> Do you guys think that pileupAsGRanges should be pushed up somewhere, >> outside of >> ggbio, or is it too high-level? There are clearly many ways that the >> pileup >> output could be coerced to GRanges; maybe Rsamtools or GenomicRanges >> could try >> to support some of the most useful? >> > > I'm ok with moving this (or some similar functionality) to Rsamtools; is > the output of pileupAsGRanges what you're interested in, or could it be > more useful? > > >> Michael >> >> >> >> >> On Mon, Dec 17, 2012 at 9:22 AM, Martin Morgan <mtmorgan@fhcrc.org>> <mailto:mtmorgan@fhcrc.org>> wrote: >> >> On 12/17/2012 03:12 AM, Mark Dunning wrote: >> >> Hi all, >> >> I'm running the pileupAsGRanges function from ggbio, and just >> wondering if there was a limit to the depth it reports? I have >> targeted sequencing data and expect many thousands of reads per >> base >> but the depth seems to be around 250? >> >> >> Hi Mark -- >> >> applyPileups takes a 'PileupParam' argument, which has maxDepth=250L >> as an >> argument, so maxDepth = .Machine$integer.max would get you everything. >> >> The ... arguments of pileupAsGRanges gets passed to PileupParam, so I >> think >> you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). >> >> Martin >> >> >> >> pgr[3:5, >> >> + ] >> GRanges with 3 ranges and 7 metadata columns: >> seqnames ranges strand | A C >> G >> <rle> <iranges> <rle> | <integer> <integer> >> <integer> >> [1] chr17 [7577489, 7577489] + | 1 0 >> 249 >> [2] chr17 [7577490, 7577490] + | 0 6 >> 245 >> [3] chr17 [7577491, 7577491] + | 1 250 >> 0 >> T N depth bam >> <integer> <integer> <numeric> <character> >> [1] 0 0 250 CleanedBams/FLD0097.bam >> [2] 0 0 251 CleanedBams/FLD0097.bam >> [3] 1 0 252 CleanedBams/FLD0097.bam >> >> >> I think that samtools mpileup has a default depth of 250, so is >> it the >> case that the applyPileup function is using the same limit, and >> it is >> possible to change it? >> >> Mark >> >> ______________________________**___________________ >> Bioconductor mailing list >> Bioconductor@r-project.org <mailto:bioconductor@r-**project.org<bioconductor@r-project.org> >> > >> https://stat.ethz.ch/mailman/_**_listinfo/bioconductor<http s:="" stat.ethz.ch="" mailman="" __listinfo="" bioconductor=""> >> >> <https: stat.ethz.ch="" mailman="" **listinfo="" bioconductor<https="" :="" stat.ethz.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=""> >> >> <http: news.gmane.org="" gmane.**science.biology.informatics.**="">> conductor<http: news.gmane.org="" gmane.science.biology.informatics.c="" onductor=""> >> > >> >> >> >> -- >> 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 <tel:%28206%29%20667-2793> >> >> >> ______________________________**___________________ >> Bioconductor mailing list >> Bioconductor@r-project.org <mailto:bioconductor@r-**project.org<bioconductor@r-project.org> >> > >> https://stat.ethz.ch/mailman/_**_listinfo/bioconductor<https: stat.ethz.ch="" mailman="" __listinfo="" bioconductor=""> >> >> <https: stat.ethz.ch="" mailman="" **listinfo="" bioconductor<https:="" s="" tat.ethz.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=""> >> <http: news.gmane.org="" gmane.**science.biology.informatics.**="">> conductor<http: news.gmane.org="" gmane.science.biology.informatics.c="" onductor=""> >> > >> >> >> > > -- > 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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Hi Martin, Thanks for the info. I managed to work it out in the end. The tip about .Machine$integer.max is useful to know! Is it possible for the function to return insertions / deletions at each position, or just base counts? Mark On Mon, Dec 17, 2012 at 5:22 PM, Martin Morgan <mtmorgan at="" fhcrc.org=""> wrote: > On 12/17/2012 03:12 AM, Mark Dunning wrote: >> >> Hi all, >> >> I'm running the pileupAsGRanges function from ggbio, and just >> wondering if there was a limit to the depth it reports? I have >> targeted sequencing data and expect many thousands of reads per base >> but the depth seems to be around 250? > > > Hi Mark -- > > applyPileups takes a 'PileupParam' argument, which has maxDepth=250L as an > argument, so maxDepth = .Machine$integer.max would get you everything. > > The ... arguments of pileupAsGRanges gets passed to PileupParam, so I think > you can add maxDepth=.Machine$integer.max to pileupAsGRanges(). > > Martin > >> >>> pgr[3:5, >> >> + ] >> GRanges with 3 ranges and 7 metadata columns: >> seqnames ranges strand | A C G >> <rle> <iranges> <rle> | <integer> <integer> <integer> >> [1] chr17 [7577489, 7577489] + | 1 0 249 >> [2] chr17 [7577490, 7577490] + | 0 6 245 >> [3] chr17 [7577491, 7577491] + | 1 250 0 >> T N depth bam >> <integer> <integer> <numeric> <character> >> [1] 0 0 250 CleanedBams/FLD0097.bam >> [2] 0 0 251 CleanedBams/FLD0097.bam >> [3] 1 0 252 CleanedBams/FLD0097.bam >> >> >> I think that samtools mpileup has a default depth of 250, so is it the >> case that the applyPileup function is using the same limit, and it is >> possible to change it? >> >> Mark >> >> _______________________________________________ >> 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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