diffHic:Fails to load bam file during preparePairs
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@manjulathimma-11250
Last seen 8.2 years ago

Hi Aaron,

Hi After removing extra chromosomes, I have loaded my bam files.

But facing the following error.

Ago1 <- preparePairs("/Users/data//HiCMapped/Ago1_presplit_out_fixmate_nochrM_new.bam",hs.param,file="/Users/thimmamp/temp_tobe_deleted/HiCMapped/Ago1_diffhic.h5",dedup=TRUE,minq=10)
Error in preparePairs("/Users/data/HiCMapped/Ago1_presplit_out_fixmate_nochrM_new.bam",  :
  failed to open output file at './file3387852bd5/17_15'

diffhic • 1.3k views
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Aaron Lun ★ 28k
@alun
Last seen 26 minutes ago
The city by the bay

What version of diffHic are you using? This seems like the bug that was caused by having too many file handles open; it should have been fixed in version 1.4.3.

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@manjulathimma-11250
Last seen 8.2 years ago

Thanks Aaron, I downloaded the latest version and this issue is fixed!

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@manjulathimma-11250
Last seen 8.2 years ago

Hi Aaron, I am trying to count interacting fragments in pre-defined regions.

But there seems to an error.

redata <- connectCounts(input, hs.param, regions=gene.body)
Error in connectCounts(input, hs.param, regions = gene.body) :
  chromosome present in regions and not in fragments
In addition: Warning message:
In connectCounts(input, hs.param, regions = gene.body) :
  stranded region ranges have no interpretation, coercing unstrandedness
> input
[1] "/Users/thimmamp/temp_tobe_deleted/HiCMapped/HepG2_trimmed.h5" "/Users/thimmamp/temp_tobe_deleted/HiCMapped/Ago1_trimmed.h5"

> sessionInfo()
R version 3.3.1 (2016-06-21)
Platform: x86_64-apple-darwin13.4.0 (64-bit)
Running under: OS X 10.9.5 (Mavericks)

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] TxDb.Hsapiens.UCSC.hg19.knownGene_3.2.2 GenomicFeatures_1.24.5                  AnnotationDbi_1.34.4                    BiocInstaller_1.22.3                    diffHic_1.4.3                          
 [6] InteractionSet_1.0.4                    SummarizedExperiment_1.2.3              Biobase_2.32.0                          GenomicRanges_1.24.2                    GenomeInfoDb_1.8.3                     
[11] IRanges_2.6.1                           S4Vectors_0.10.2                        BiocGenerics_0.18.0                    

loaded via a namespace (and not attached):
 [1] edgeR_3.14.0            XVector_0.12.1          zlibbioc_1.18.0         GenomicAlignments_1.8.4 BiocParallel_1.6.3      BSgenome_1.40.1         lattice_0.20-33         tools_3.3.1            
 [9] grid_3.3.1              rhdf5_2.16.0            csaw_1.6.1              DBI_0.4-1               Rhtslib_1.4.3           Matrix_1.2-6            rtracklayer_1.32.2      bitops_1.0-6           
[17] biomaRt_2.28.0          RCurl_1.95-4.8          RSQLite_1.0.0           limma_3.28.17           Rsamtools_1.24.0        Biostrings_2.40.2       locfit_1.5-9.1          XML_3.98-1.4           
 

Would you please help me resolve this?

 

 

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This probably should go into a new question, but I'll answer it here. The problem is that you've got some intervals in regions that are on chromosomes that don't exist in hs.param$fragmentsdiffHic isn't quite sure what to do with those regions, hence it throws an error. The solution is to screen out these regions before supplying them to connectCounts. I guess the function could be more forgiving and do this automatically.

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