[Bioc-devel] error using the DEXSeqDataSet function

Martin Morgan mtmorgan at fredhutch.org
Thu Jul 30 19:59:43 CEST 2015


On 07/30/2015 09:02 AM, Alejandro Reyes wrote:
> Dear Leonard,
>
> Thanks a lot for reporting this. It should be fixed in the version that
> I just committed to the svn (DEXSeq 1.5.10).
>
> While debugging the DEXSeq code, I noticed that summarizedOverlaps is
> giving me an error, which I think its a bug while creating the
> summarizedExperiments object that is returned. Here a reproducible example:
>
> Konsole output
>>     library(GenomicRanges)
>   >   library(GenomicFeatures)
>   >   library(GenomicAlignments)
>   >
>   >   hse <- makeTxDbFromBiomart( biomart="ensembl",
> +                               dataset="hsapiens_gene_ensembl" )
>
>
> Download and preprocess the 'transcripts' data frame ... OK
> Download and preprocess the 'chrominfo' data frame ... OK
> Download and preprocess the 'splicings' data frame ... OK
> Download and preprocess the 'genes' data frame ... OK
> Prepare the 'metadata' data frame ... OK
> Make the TxDb object ... OK
>   >
>   >   bamDir <- system.file(
> +     "extdata", package="parathyroidSE", mustWork=TRUE )
>   >   fls <- list.files( bamDir, pattern="bam$", full=TRUE )
>   >
>   >   bamlst <- BamFileList(
> +     fls, index=character(),
> +     yieldSize=100000, obeyQname=TRUE )
>   >
>   >   exonicParts <- disjointExons( hse, aggregateGenes=FALSE )
>   >
>   >   SE <- summarizeOverlaps( exonicParts, bamlst,
> +     mode="Union", singleEnd=FALSE,
> +     ignore.strand=TRUE, inter.feature=FALSE, fragments=TRUE )
> Error in SummarizedExperiment(assays = SimpleList(counts = counts),
> rowRanges = features,  :
>    error in evaluating the argument 'assays' in selecting a method for
> function 'SummarizedExperiment': Error in validObject(.Object) :
>    invalid class �SimpleList� object: invalid object for slot "listData"
> in class "SimpleList": got class "matrix", should be or extend class "lis
> t"

with options(error=recover) we end up at

Enter a frame number, or 0 to exit

  1: summarizeOverlaps(exonicParts, bamlst, mode = "Union", singleEnd = FALSE, i
  2: summarizeOverlaps(exonicParts, bamlst, mode = "Union", singleEnd = FALSE, i
  3: .local(features, reads, mode, algorithm, ignore.strand, ...)
  4: .dispatchBamFiles(features, reads, mode, match.arg(algorithm), ignore.stran
  5: SummarizedExperiment(assays = SimpleList(counts = counts), rowRanges = feat
  6: SimpleList(counts = counts)
  7: new("SimpleList", listData = args)
  8: initialize(value, ...)
  9: initialize(value, ...)
10: validObject(.Object)

Selection: 4
Called from: stop(msg, ": ", errors, domain = NA)
Browse[2]> SimpleList(counts=counts)
Error during wrapup: invalid class "SimpleList" object: invalid object for slot 
"listData" in class "SimpleList": got class "matrix", should be or extend class 
"list"
Browse[2]> class(counts)
[1] "matrix"
Browse[2]> dim(counts)  ## suspicious!
[1] 2 3
Browse[2]> cat(counts[[1]])
sequences 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 
21, 22, X, Y have incompatible seqlengths:
   - in 'x': 248956422, 242193529, 198295559, 190214555, 181538259, 170805979, 
159345973, 145138636, 138394717, 133797422, 135086622, 133275309, 114364328, 
107043718, 101991189, 90338345, 83257441, 80373285, 58617616, 64444167, 
46709983, 50818468, 156040895, 57227415
   - in 'y': 249250621, 243199373, 198022430, 191154276, 180915260, 171115067, 
159138663, 146364022, 141213431, 135534747, 135006516, 133851895, 115169878, 
107349540, 102531392, 90354753, 81195210, 78077248, 59128983, 63025520, 
48129895, 51304566, 155270560, 59373566

So (a) the genome build of the alignment is different from the genome build of 
the annotations (b) BiocParallel (I think) is capturing the messages rather than 
the return values (c) SimpleList(matrix(list())) fails.

Martin


>
> And the output of sessionInfo(),
>
> Konsole output
>>   sessionInfo()
> R Under development (unstable) (2015-07-25 r68744)
> Platform: x86_64-pc-linux-gnu (64-bit)
> Running under: Ubuntu 15.04
>
> locale:
> [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C
> [3] LC_TIME=de_DE.UTF-8        LC_COLLATE=en_US.UTF-8
> [5] LC_MONETARY=de_DE.UTF-8    LC_MESSAGES=en_US.UTF-8
> [7] LC_PAPER=de_DE.UTF-8       LC_NAME=C
> [9] LC_ADDRESS=C               LC_TELEPHONE=C
> [11] LC_MEASUREMENT=de_DE.UTF-8 LC_IDENTIFICATION=C
>
> attached base packages:
> [1] stats4    parallel  stats     graphics  grDevices utils     datasets
> [8] methods   base
>
> other attached packages:
> [1] GenomicAlignments_1.5.12   Rsamtools_1.21.14
> [3] Biostrings_2.37.2          XVector_0.9.1
> [5] GenomicFeatures_1.21.13    AnnotationDbi_1.31.17
> [7] DEXSeq_1.15.10             DESeq2_1.9.26
> [9] RcppArmadillo_0.5.200.1.0  Rcpp_0.12.0
> [11] SummarizedExperiment_0.3.2 GenomicRanges_1.21.17
> [13] GenomeInfoDb_1.5.9         IRanges_2.3.15
> [15] S4Vectors_0.7.10           Biobase_2.29.1
> [17] BiocGenerics_0.15.3        BiocParallel_1.3.42
>
> loaded via a namespace (and not attached):
> [1] genefilter_1.51.0    statmod_1.4.21       locfit_1.5-9.1
> [4] reshape2_1.4.1       splines_3.3.0        lattice_0.20-33
> [7] colorspace_1.2-6     rtracklayer_1.29.12  survival_2.38-3
> [10] XML_3.98-1.3         foreign_0.8-65       DBI_0.3.1
> [13] RColorBrewer_1.1-2   lambda.r_1.1.7       plyr_1.8.3
> [16] stringr_1.0.0        zlibbioc_1.15.0      munsell_0.4.2
> [19] gtable_0.1.2         futile.logger_1.4.1  hwriter_1.3.2
> [22] latticeExtra_0.6-26  geneplotter_1.47.0   biomaRt_2.25.1
> [25] proto_0.3-10         acepack_1.3-3.3      xtable_1.7-4
> [28] scales_0.2.5         Hmisc_3.16-0         annotate_1.47.4
> [31] gridExtra_2.0.0      ggplot2_1.0.1        digest_0.6.8
> [34] stringi_0.5-5        grid_3.3.0           tools_3.3.0
> [37] bitops_1.0-6         magrittr_1.5         RCurl_1.95-4.7
> [40] RSQLite_1.0.0        Formula_1.2-1        cluster_2.0.3
> [43] futile.options_1.0.0 MASS_7.3-43          rpart_4.1-10
> [46] nnet_7.3-10
>
>
> Best regards,
> Alejandro Reyes
>
>
>
>
> On 29.07.2015 20:26, Leonard Goldstein wrote:
>> Hi all,
>>
>> I'm having trouble creating a DEXSeqDataSet object (in the devel
>> version of DEXSeq)
>>
>> Running the example included in the manual page results in the same
>> error I get with my own data (see below)
>>
>> Many thanks for your help.
>>
>> Leonard
>>
>>> library(DEXSeq)
>>> countData <- matrix( rpois(10000, 100), nrow=1000 )
>>> sampleData <- data.frame(
>> +     condition=rep( c("untreated", "treated"), each=5 ) )
>>> design <- formula( ~ sample + exon + condition:exon )
>>> groupID <- rep(
>> +     paste0("gene", 1:10),
>> +     each= 100 )
>>> featureID <- rep(
>> +     paste0("exon", 1:10),
>> +     times= 100 )
>>> DEXSeqDataSet( countData, sampleData, design,
>> +             featureID, groupID )
>> converting counts to integer mode
>>
>> Error in `$<-.data.frame`(`*tmp*`, "dispersion", value = NA) :
>>     replacement has 1 row, data has 0
>> In addition: Warning message:
>> In DESeqDataSet(se, design, ignoreRank = TRUE) :
>>     900 duplicate rownames were renamed by adding numbers
>>
>>> sessionInfo()
>> R version 3.2.1 (2015-06-18)
>> Platform: x86_64-unknown-linux-gnu (64-bit)
>> Running under: Red Hat Enterprise Linux Server release 6.6 (Santiago)
>>
>> 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] stats4    parallel  stats     graphics  grDevices utils     datasets
>> [8] methods   base
>>
>> other attached packages:
>>    [1] DEXSeq_1.15.9              DESeq2_1.9.26
>>    [3] RcppArmadillo_0.5.200.1.0  Rcpp_0.12.0
>>    [5] SummarizedExperiment_0.3.2 GenomicRanges_1.21.17
>>    [7] GenomeInfoDb_1.5.9         IRanges_2.3.15
>>    [9] S4Vectors_0.7.10           Biobase_2.29.1
>> [11] BiocGenerics_0.15.3        BiocParallel_1.3.41
>>
>> loaded via a namespace (and not attached):
>>    [1] genefilter_1.51.0     statmod_1.4.21        locfit_1.5-9.1
>>    [4] reshape2_1.4.1        splines_3.2.1         lattice_0.20-33
>>    [7] colorspace_1.2-6      survival_2.38-3       XML_3.98-1.3
>> [10] foreign_0.8-65        DBI_0.3.1             RColorBrewer_1.1-2
>> [13] lambda.r_1.1.7        plyr_1.8.3            stringr_1.0.0
>> [16] zlibbioc_1.15.0       Biostrings_2.37.2     munsell_0.4.2
>> [19] gtable_0.1.2          futile.logger_1.4.1   hwriter_1.3.2
>> [22] latticeExtra_0.6-26   geneplotter_1.47.0    biomaRt_2.25.1
>> [25] AnnotationDbi_1.31.17 proto_0.3-10          acepack_1.3-3.3
>> [28] xtable_1.7-4          scales_0.2.5          Hmisc_3.16-0
>> [31] annotate_1.47.4       XVector_0.9.1         Rsamtools_1.21.14
>> [34] gridExtra_2.0.0       ggplot2_1.0.1         digest_0.6.8
>> [37] stringi_0.5-5         grid_3.2.1            tools_3.2.1
>> [40] bitops_1.0-6          magrittr_1.5          RCurl_1.95-4.7
>> [43] RSQLite_1.0.0         Formula_1.2-1         cluster_2.0.3
>> [46] futile.options_1.0.0  MASS_7.3-43           rpart_4.1-10
>> [49] compiler_3.2.1        nnet_7.3-10
>> _______________________________________________
>> Bioc-devel at r-project.org mailing list
>> https://stat.ethz.ch/mailman/listinfo/bioc-devel
>
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