[Bioc-devel] cigarToRleList fails

Martin Morgan mtmorgan at fhcrc.org
Thu Feb 20 23:55:59 CET 2014


On 02/20/2014 02:32 PM, Hervé Pagès wrote:
> Hi Jesper,
>
> On 02/20/2014 02:13 PM, Jesper Gådin wrote:
>> Very true that it is quite difficult to find the documentation when one
>> is not aware of its existence :P
>
> Yeah, this has been a source of frustration for many people. And
> always a source of embarrassment (for us) when teaching our software
> to beginners.
>
> I've started to change this. In the upcoming version of BioC (2.14,
> scheduled for mid-April), when you'll do ?coverage, you'll get to
> choose between the 3 man pages that document coverage methods (there
> is one in IRanges, one in GenomicRanges, and one in GenomicAlignments).
>
> I want to generalize this to other generics that have methods spread
> across several packages (e.g. findOverlaps, the intra- and inter-range
> methods, etc...).
>
> Having to choose between several man pages every time you do e.g.
> ?findOverlaps is a minor annoyance compared to not being able to
> find the man page at all. (Note that if you already know where is
> your favorite man page, you'll be able to direct access it with
> e.g. ?GenomicRanges::findOverlaps). Nobody will ever need to use
> the insane ?`findOverlaps,GenomicRanges,GenomicRanges-method` to

tab completion helps, so that you don't need to be totally insane, just insane 
enough to know to start with

?"cover<tab>

Martin

> open that man page again. Ever! (it will still work though...)
>
> Cheers,
> H.
>
>>
>> coverage() is fast and beautiful. Thanks!
>>
>> /Jesper
>>
>>
>> On Wed, Feb 19, 2014 at 9:21 PM, Hervé Pagès <hpages at fhcrc.org
>> <mailto:hpages at fhcrc.org>> wrote:
>>
>>     Hi Jesper,
>>
>>
>>     On 02/19/2014 08:44 AM, Michael Lawrence wrote:
>>
>>         On Wed, Feb 19, 2014 at 8:39 AM, Jesper Gådin
>>         <jesper.gadin at gmail.com <mailto:jesper.gadin at gmail.com>>wrote:
>>
>>             Hi Michael,
>>
>>             Herves suggestion will probably work for my use case, but if
>>             there are any
>>             smoother ways it would be preferable.
>>
>>             The use case is as follows:
>>
>>             1) calculate strand specific coverage over a region from
>>             GAlignments object (or file)
>>
>>             At the moment I read a file using readGAlignmentsFromBam()
>>             with tag="XS",
>>             then filter it on "flag" and "mapq". Then I subset the
>>             resulting GAlignments in a minus and a plus -strand object.
>>             Then I calculate coverage by my own coverage function which
>>             uses the cigar
>>             information in the GAlignments object. This function is the
>>             one using
>>             cigarToRleList() at the moment. As I understand the
>>             coverage() function
>>             from the GenomicAlignments/IRanges package does not take
>>             into account
>>             cigars, or does it?
>>
>>
>>         It does take the coverage into account; specifically to exclude
>>         the introns
>>         from coverage. I think there's also an option to exclude deletions.
>>
>>
>>     Unfortunately the man page is not easy to access (you need to do
>>     ?`coverage,GAlignments-method`__), but it says:
>>
>>         The methods for GAlignments and GAlignmentPairs objects do:
>>
>>           coverage(grglist(x), ...)
>>
>>     And if you do grglist() on a GAlignments or GAlignmentPairs objects, the
>>     ranges you get in the returned GRangesList object are calculated based
>>     on the CIGAR.
>>
>>     Trust but verify. Here is how you can actually verify that coverage()
>>     does take the CIGAR into account:
>>
>>        library(RNAseqData.HNRNPC.bam.__chr14)
>>        gal <- readGAlignments(RNAseqData.__HNRNPC.bam.chr14_BAMFILES[1])
>>        cig_op_table <- cigarOpTable(cigar(gal))
>>
>>     First we pick up an alignment with an N in its CIGAR and do coverage()
>>     on it:
>>
>>        > gal_with_N <- gal[which(cig_op_table[ , "N"] != 0)[1]]
>>
>>        > gal_with_N
>>        GAlignments with 1 alignment and 0 metadata columns:
>>              seqnames strand       cigar    qwidth     start       end
>>        width
>>                 <Rle>  <Rle> <character> <integer> <integer> <integer>
>>     <integer>
>>          [1]    chr14      + 55M2117N17M        72  19650072  19652260
>>         2189
>>                   ngap
>>              <integer>
>>          [1]         1
>>          ---
>>          seqlengths:
>>                          chr1                 chr10 ...
>>       chrY
>>                     249250621             135534747 ...
>>       59373566
>>
>>        > coverage(gal_with_N)$chr14
>>        integer-Rle of length 107349540 with 5 runs
>>          Lengths: 19650071       55     2117       17 87697280
>>          Values :        0        1        0        1        0
>>
>>     Same thing with an alignment with an I in its CIGAR:
>>
>>        > gal_with_I <- gal[which(cig_op_table[ , "I"] != 0)[1]]
>>
>>        > gal_with_I
>>        GAlignments with 1 alignment and 0 metadata columns:
>>              seqnames strand       cigar    qwidth     start       end
>>        width
>>                 <Rle>  <Rle> <character> <integer> <integer> <integer>
>>     <integer>
>>          [1]    chr14      -     64M1I7M        72  19411677  19411747
>>           71
>>                   ngap
>>              <integer>
>>          [1]         0
>>          ---
>>          seqlengths:
>>                          chr1                 chr10 ...
>>       chrY
>>                     249250621             135534747 ...
>>       59373566
>>
>>        > coverage(gal_with_I)$chr14
>>        integer-Rle of length 107349540 with 3 runs
>>          Lengths: 19411676 71 87937793 <tel:71%2087937793>
>>          Values :        0        1        0
>>
>>     Same thing with an alignment with a D in its CIGAR:
>>
>>        > gal_with_D <- gal[which(cig_op_table[ , "D"] != 0)[1]]
>>
>>        > gal_with_D
>>        GAlignments with 1 alignment and 0 metadata columns:
>>              seqnames strand       cigar    qwidth     start       end
>>        width
>>                 <Rle>  <Rle> <character> <integer> <integer> <integer>
>>     <integer>
>>          [1]    chr14      +    38M1D34M        72  19659063  19659135
>>           73
>>                   ngap
>>              <integer>
>>          [1]         0
>>          ---
>>          seqlengths:
>>                          chr1                 chr10 ...
>>       chrY
>>                     249250621             135534747 ...
>>       59373566
>>
>>        > coverage(gal_with_D)$chr14
>>        integer-Rle of length 107349540 with 3 runs
>>          Lengths: 19659062       73 87690405
>>          Values :        0        1        0
>>
>>     Seeing is believing,
>>
>>     Cheers,
>>     H.
>>
>>
>>
>>             I started to look at the applyPileups() in Rsamtools which I
>>             can get to
>>             calculate coverage using cigars, but not using the strand or
>>             flag
>>             information for filtering. That solution would start from a
>>             bam-file
>>             instead of a GAlignments object, and sure I can do the
>>             filtering outside R.
>>             But it would be very convenient to do it all from within R.
>>
>>             If there are nice solutions starting from both a GAlignments
>>             and a
>>             bam-file it would be great! =)
>>
>>             /Jesper
>>
>>
>>
>>             On Tue, Feb 18, 2014 at 10:52 PM, Michael Lawrence <
>>             lawrence.michael at gene.com
>>             <mailto:lawrence.michael at gene.com>> wrote:
>>
>>                 Hi Jesper,
>>
>>                 Would you be willing to volunteer your use case? As
>>                 Herve hinted,
>>                 cigarToRleList and friends are low-level helpers. There
>>                 may be an easier
>>                 way to achieve what you want, or an opportunity to
>>                 improve things.
>>
>>                 Michael
>>
>>
>>                 On Mon, Feb 17, 2014 at 1:10 AM, Jesper Gådin
>>                 <jesper.gadin at gmail.com
>>                 <mailto:jesper.gadin at gmail.com>>wrote:
>>
>>                     Hi,
>>
>>                     Have come across a cigar-vector that is problematic
>>                     to process.
>>
>>                     #load package
>>
>>                         library(GenomicAlignments)
>>
>>
>>                     #load data (see attached file)
>>
>>                         load("2014-02-17-cigarExample.__rdata")
>>
>>
>>                     #run function cigarToRleList
>>
>>                         cigarRle <- cigarToRleList(cigarExample)
>>
>>                     Error in .Call2("Rle_constructor", values, lengths,
>>                     check, 0L, PACKAGE =
>>                     "IRanges") :
>>                         integer overflow while summing elements in 'lengths'
>>
>>                         sessionInfo()
>>
>>                     R Under development (unstable) (2013-11-13 r64209)
>>                     Platform: x86_64-unknown-linux-gnu (64-bit)
>>
>>                     locale:
>>                        [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C
>>                        [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8
>>                        [5] LC_MONETARY=en_US.UTF-8
>>                       LC_MESSAGES=en_US.UTF-8
>>                        [7] LC_PAPER=en_US.UTF-8       LC_NAME=C
>>                        [9] LC_ADDRESS=C               LC_TELEPHONE=C
>>                     [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C
>>
>>                     attached base packages:
>>                     [1] parallel  stats     graphics  grDevices utils
>>                        datasets  methods
>>                     [8] base
>>
>>                     other attached packages:
>>                     [1] GenomicAlignments_0.99.18 Rsamtools_1.15.26
>>                     [3] Biostrings_2.31.12        XVector_0.3.6
>>                     [5] GenomicRanges_1.15.26     IRanges_1.21.23
>>                     [7] BiocGenerics_0.9.3
>>
>>                     loaded via a namespace (and not attached):
>>                        [1] BatchJobs_1.1-1135 BBmisc_1.5
>>                     BiocParallel_0.5.8
>>                     bitops_1.0-6
>>
>>                        [5] brew_1.0-6         codetools_0.2-8    DBI_0.2-7
>>                        digest_0.6.4
>>
>>                        [9] fail_1.2           foreach_1.4.1
>>                       iterators_1.0.6    plyr_1.8
>>
>>                     [13] RSQLite_0.11.4     sendmailR_1.1-2
>>                       stats4_3.1.0       tools_3.1.0
>>
>>                     [17] zlibbioc_1.9.0
>>
>>                     _________________________________________________
>>                     Bioc-devel at r-project.org
>>                     <mailto:Bioc-devel at r-project.org> mailing list
>>                     https://stat.ethz.ch/mailman/__listinfo/bioc-devel
>>                     <https://stat.ethz.ch/mailman/listinfo/bioc-devel>
>>
>>
>>
>>
>>
>>                  [[alternative HTML version deleted]]
>>
>>
>>
>>
>>         _________________________________________________
>>         Bioc-devel at r-project.org <mailto:Bioc-devel at r-project.org>
>>         mailing list
>>         https://stat.ethz.ch/mailman/__listinfo/bioc-devel
>>         <https://stat.ethz.ch/mailman/listinfo/bioc-devel>
>>
>>
>>     --
>>     Hervé Pagès
>>
>>     Program in Computational Biology
>>     Division of Public Health Sciences
>>     Fred Hutchinson Cancer Research Center
>>     1100 Fairview Ave. N, M1-B514
>>     P.O. Box 19024
>>     Seattle, WA 98109-1024
>>
>>     E-mail: hpages at fhcrc.org <mailto:hpages at fhcrc.org>
>>     Phone: (206) 667-5791 <tel:%28206%29%20667-5791>
>>     Fax: (206) 667-1319 <tel:%28206%29%20667-1319>
>>
>>
>


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