[Bioc-devel] BamTallyParam argument 'which'

Leonardo Collado Torres lcollado at jhu.edu
Wed Feb 25 04:40:12 CET 2015


Related to my post on a separate thread
(https://stat.ethz.ch/pipermail/bioc-devel/2015-February/006978.html),
I think that if 'which' is not being reduced by default, a simple
example showing the effects of this could be included in the functions
that have such an argument. Also note that 'reducing' could lead to
unintended results.

For example, in the help page for GenomicAlignments::readGAlignments,
after the 'gal4' example it would be nice to add something like this:


## Note that if overlapping ranges are provided in 'which'
## reads could be selected more than once. This would artificually
## increase the coverage or affect other downstream results.
## If you 'reduce' the ranges, reads that originally overlapped
## two disjoint segments will be included.

which_dups <- RangesList(seq1=rep(IRanges(1000, 2000), 2),
    seq2=IRanges(c(100, 1000), c(1000, 2000)))
param_dups <- ScanBamParam(which=which_dups)
param_reduced <- ScanBamParam(which=reduce(which_dups))
gal4_dups <- readGAlignments(bamfile, param=param_dups)
gal4_reduced <- readGAlignments(bamfile, param=param_reduced)


length(gal4)

## Duplicates some reads. In this case, all the ones between
## bases 1000 and 2000 on seq1.
length(gal4_dups)

## Includes some reads that mapped around base 1000 in seq2
## that were excluded in gal4.
length(gal4_reduced)








Here's the output:



> library('GenomicAlignments')
>
> ## Code already included in ?readGAlignments
> bamfile <- system.file("extdata", "ex1.bam", package="Rsamtools",
+                        mustWork=TRUE)
> which <- RangesList(seq1=IRanges(1000, 2000),
+                     seq2=IRanges(c(100, 1000), c(1000, 2000)))
> param <- ScanBamParam(which=which)
> gal4 <- readGAlignments(bamfile, param=param)
> gal4
GAlignments object with 2404 alignments and 0 metadata columns:
         seqnames strand       cigar    qwidth     start       end
width     njunc
            <Rle>  <Rle> <character> <integer> <integer> <integer>
<integer> <integer>
     [1]     seq1      +         35M        35       970      1004
   35         0
     [2]     seq1      +         35M        35       971      1005
   35         0
     [3]     seq1      +         35M        35       972      1006
   35         0
     [4]     seq1      +         35M        35       973      1007
   35         0
     [5]     seq1      +         35M        35       974      1008
   35         0
     ...      ...    ...         ...       ...       ...       ...
  ...       ...
  [2400]     seq2      +         35M        35      1524      1558
   35         0
  [2401]     seq2      +         35M        35      1524      1558
   35         0
  [2402]     seq2      -         35M        35      1528      1562
   35         0
  [2403]     seq2      -         35M        35      1532      1566
   35         0
  [2404]     seq2      -         35M        35      1533      1567
   35         0
  -------
  seqinfo: 2 sequences from an unspecified genome
>
> ## Note that if overlapping ranges are provided in 'which'
> ## reads could be selected more than once. This would artificually
> ## increase the coverage or affect other downstream results.
> ## If you 'reduce' the ranges, reads that originally overlapped
> ## two disjoint segments will be included.
>
> which_dups <- RangesList(seq1=rep(IRanges(1000, 2000), 2),
+     seq2=IRanges(c(100, 1000), c(1000, 2000)))
> param_dups <- ScanBamParam(which=which_dups)
> param_reduced <- ScanBamParam(which=reduce(which_dups))
> gal4_dups <- readGAlignments(bamfile, param=param_dups)
> gal4_reduced <- readGAlignments(bamfile, param=param_reduced)
>
>
> length(gal4)
[1] 2404
>
> ## Duplicates some reads. In this case, all the ones between
> ## bases 1000 and 2000 on seq1.
> length(gal4_dups)
[1] 3014
>
> ## Includes some reads that mapped around base 1000 in seq2
> ## that were excluded in gal4.
> length(gal4_reduced)
[1] 2343
>
>
>
>
>
> options(width = 120)
> devtools::session_info()
Session info-----------------------------------------------------------------------------------------------------------
 setting  value
 version  R Under development (unstable) (2014-11-01 r66923)
 system   x86_64, darwin10.8.0
 ui       AQUA
 language (EN)
 collate  en_US.UTF-8
 tz       America/New_York

Packages---------------------------------------------------------------------------------------------------------------
 package           * version date       source
 base64enc           0.1.2   2014-06-26 CRAN (R 3.2.0)
 BatchJobs           1.4     2014-09-24 CRAN (R 3.2.0)
 BBmisc              1.7     2014-06-21 CRAN (R 3.2.0)
 BiocGenerics      * 0.13.4  2014-12-31 Bioconductor
 BiocParallel        1.1.13  2015-01-27 Bioconductor
 Biostrings        * 2.35.8  2015-02-14 Bioconductor
 bitops              1.0.6   2013-08-17 CRAN (R 3.2.0)
 brew                1.0.6   2011-04-13 CRAN (R 3.2.0)
 checkmate           1.5.0   2014-10-19 CRAN (R 3.2.0)
 codetools           0.2.9   2014-08-21 CRAN (R 3.2.0)
 DBI                 0.3.1   2014-09-24 CRAN (R 3.2.0)
 devtools            1.6.1   2014-10-07 CRAN (R 3.2.0)
 digest              0.6.4   2013-12-03 CRAN (R 3.2.0)
 fail                1.2     2013-09-19 CRAN (R 3.2.0)
 foreach             1.4.2   2014-04-11 CRAN (R 3.2.0)
 GenomeInfoDb      * 1.3.13  2015-02-13 Bioconductor
 GenomicAlignments * 1.3.27  2015-01-26 Bioconductor
 GenomicRanges     * 1.19.37 2015-02-13 Bioconductor
 IRanges           * 2.1.38  2015-02-08 Bioconductor
 iterators           1.0.7   2014-04-11 CRAN (R 3.2.0)
 Rsamtools         * 1.19.27 2015-02-07 Bioconductor
 RSQLite             1.0.0   2014-10-25 CRAN (R 3.2.0)
 rstudioapi          0.1     2014-03-27 CRAN (R 3.2.0)
 S4Vectors         * 0.5.20  2015-02-19 Bioconductor
 sendmailR           1.2.1   2014-09-21 CRAN (R 3.2.0)
 stringr             0.6.2   2012-12-06 CRAN (R 3.2.0)
 XVector           * 0.7.4   2015-02-08 Bioconductor
 zlibbioc            1.13.1  2015-02-11 Bioconductor
>





On Mon, Feb 23, 2015 at 2:38 PM, Leonard Goldstein
<goldstein.leonard at gene.com> wrote:
> Sounds very sensible not to double count in the context of tallying
> variants. I was more concerned with reducing which as the default
> behavior for scanBam and other functions.
>
> I wanted to bring up the samtools behavior as - for me at least -
> inconsistencies between Rsamtools and samtools have been another
> source of confusion in the past (e.g. different naming conventions for
> fields like isize vs TLEN etc.)
>
> Leonard
>
>
> On Mon, Feb 23, 2015 at 11:22 AM, Michael Lawrence
> <lawrence.michael at gene.com> wrote:
>> Maybe Rsamtools would want to follow this precedent. I think there might be
>> a difference between fishing out alignments from a SAM/BAM, and deriving a
>> summary (tallyVariants) from a BAM. It seems like an argument could be made
>> for a tally set to not contain duplicates.
>>
>> On Mon, Feb 23, 2015 at 11:05 AM, Leonard Goldstein
>> <goldstein.leonard at gene.com> wrote:
>>>
>>> Hi Michael and Thomas,
>>>
>>> I ran into the same problem in the past (i.e. when I started working
>>> with functions like scanBam I expected them not to return the same
>>> alignment multiple times)
>>>
>>> One thing to consider might be that returning alignments multiple
>>> times is consistent with the behavior of the samtools view command.
>>> Quoting from the samtools manual:
>>>
>>> “Important note: when multiple regions are given, some alignments may
>>> be output multiple times if they overlap more than one of the
>>> specified regions.”
>>>
>>> Maybe there is an argument for keeping things consistent with
>>> samtools? As you said, if documented properly, the user can decide
>>> whether to reduce regions specified in which or not.
>>>
>>> Leonard
>>>
>>>
>>> On Mon, Feb 23, 2015 at 10:52 AM, Michael Lawrence
>>> <lawrence.michael at gene.com> wrote:
>>> > We should at leaast try to avoid surprising the user. Seems like most
>>> > people expect "which" to be a simple restriction, so I think for now I
>>> > will
>>> > just reduce the which, and if someone has a use case for separate
>>> > queries,
>>> > we can address it in the future.
>>> >
>>> > On Mon, Feb 23, 2015 at 10:41 AM, Thomas Sandmann
>>> > <sandmann.thomas at gene.com>
>>> > wrote:
>>> >
>>> >> Personally, I don't have a use case with "meaningful loci" worth
>>> >> tracking,
>>> >> so keeping it simple would work for me.
>>> >>
>>> >> Incidentally, would it be good to deal with the 'which' parameter in a
>>> >> consistent way across different methods ? I just saw this recent post
>>> >> on
>>> >> the mailing list in which a used got confused by duplicate counts
>>> >> returned
>>> >> after passing 'which' to scanBamParam:
>>> >>
>>> >> https://stat.ethz.ch/pipermail/bioc-devel/2015-February/006978.html
>>> >>
>>> >>
>>> >> ---
>>> >>
>>> >> Thomas Sandmann, PhD
>>> >> Computational biologist
>>> >>
>>> >> Genentech, Inc.
>>> >> 1 DNA Way
>>> >> South San Francisco, CA 94080
>>> >> USA
>>> >>
>>> >> Phone: +1 650 225 6273
>>> >> Fax: +1 650 225 5389
>>> >> Email: sandmann.thomas at gene.com
>>> >>
>>> >> "If a man will begin with certainties, he shall end in doubts; but if
>>> >> he
>>> >> will be content to begin with doubts he shall end in certainties." --
>>> >> Sir
>>> >> Francis Bacon
>>> >>
>>> >>
>>> >> On Mon, Feb 23, 2015 at 10:37 AM, Michael Lawrence <
>>> >> lawrence.michael at gene.com> wrote:
>>> >>
>>> >>> We just have to decide which is the more useful interpretation of
>>> >>> which
>>> >>> -- as a simple restriction, or as a vector of meaningful locii, which
>>> >>> will
>>> >>> be analyzed individually? I would actually favor the first one (the
>>> >>> same as
>>> >>> yours), just because it's simpler. To keep track of the query ranges,
>>> >>> we
>>> >>> would need to add a new column to the returned object, which will more
>>> >>> often than not just be clutter. I guess we could introduce a new
>>> >>> parameter,
>>> >>> "reduceWhich" which defaults to TRUE and reduces the which. If FALSE,
>>> >>> it
>>> >>> instead adds the column mapping back to the original which ranges.
>>> >>>
>>> >>>
>>> >>> On Sun, Feb 22, 2015 at 2:36 PM, Thomas Sandmann <
>>> >>> sandmann.thomas at gene.com> wrote:
>>> >>>
>>> >>>> Hi Michael,
>>> >>>>
>>> >>>> ah, I see. I hadn't realized that returning the pileups separately
>>> >>>> for
>>> >>>> each region could be a desired feature, but that makes sense. I
>>> >>>> agree, as
>>> >>>> it is easy for the user to 'reduce' the ranges beforehand your first
>>> >>>> option
>>> >>>> (e.g. returning the ID of the range) would be more flexible.
>>> >>>>
>>> >>>> Perhaps you would consider adding a sentence to the documentation of
>>> >>>> 'which' on BamTallyParam's help page explaining that users might want
>>> >>>> to
>>> >>>> 'reduce' their ranges beforehand if they are only interested in a
>>> >>>> single
>>> >>>> tally for each base ?
>>> >>>>
>>> >>>> Thanks a lot !
>>> >>>> Thomas
>>> >>>>
>>> >>>>
>>> >>>
>>> >>
>>> >
>>> >         [[alternative HTML version deleted]]
>>> >
>>> > _______________________________________________
>>> > Bioc-devel at r-project.org mailing list
>>> > https://stat.ethz.ch/mailman/listinfo/bioc-devel
>>
>>
>
> _______________________________________________
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> https://stat.ethz.ch/mailman/listinfo/bioc-devel



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