Question

Stuck at "Read counting step", "summarizeOverlaps"

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JunLVI ▴ 40

@junlvi-8996

Last seen 5.7 years ago

Japan

Hi, I was following the workflow (http://www.bioconductor.org/help/workflows/rnaseqGene/)

somehow I got stuck at "Read counting step".

sampleTable <- read.csv("my_sampletable.csv",row.names=1)
filenames <- paste0(sampleTable$samplename,".bam")
file.exists(filenames)
library("Rsamtools")
bamfiles <- BamFileList(filenames, yieldSize=2000000)
gtffile <- ("mm10genes.gtf") 
library("GenomicFeatures")
(txdb <- makeTxDbFromGFF(gtffile, format="gtf", circ_seqs=character()))
(ebg <- exonsBy(txdb, by="gene"))
library("GenomicAlignments")
library("BiocParallel")
register(MulticoreParam()) # for the first try, I used "register(SerialParam())"

se <- summarizeOverlaps(features=ebg, reads=bamfiles,
                        mode="Union",
                        singleEnd=FALSE,
                        ignore.strand=TRUE,
                        fragments=TRUE )

Then, I was stuck at this step.

Any suggestion about what might go wrong?

if any information is needed for the trouble shooting, please let me know.

TKS!

deseq2 • 1.3k views

ADD COMMENT • link updated 7.3 years ago by Ryan C. Thompson ★ 7.9k • written 7.3 years ago by JunLVI ▴ 40

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What do you mean by "stuck"? Was there an error message? If you don't explain what problem you're having, there's not much we can do to help you.

ADD REPLY • link 7.3 years ago Ryan C. Thompson ★ 7.9k

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Sorry Ryan, I am sorry that I did not explain clearly.
By "stuck", I meant
when I tried to create the SummarizedExperiment object with counts:
se <- summarizeOverlaps(features=ebg, reads=bamfiles,
                        mode="Union",
                        singleEnd=FALSE,
                        ignore.strand=TRUE,
                        fragments=TRUE )

Then the computer seemed to be running the call like forever. No error message. If i wait long enough (like overnight), I got broken pipe. I wish that explained.

ADD REPLY • link 7.3 years ago JunLVI ▴ 40

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Can you (and Ryan) provide the output of the R command sessionInfo(). And also try a simple test of MulticoreParam(). I did the following

> library(BiocParallel)
> param = MulticoreParam()
> param
class: MulticoreParam
  bpisup: FALSE; bpnworkers: 6; bptasks: 0; bpjobname: BPJOB
  bplog: FALSE; bpthreshold: INFO; bpstopOnError: TRUE
  bptimeout: 2592000; bpprogressbar: FALSE
  bpRNGseed: 
  bplogdir: NA
  bpresultdir: NA
  cluster type: FORK
> bplapply(1:5, function(i) i, BPPARAM=param)
[[1]]
[1] 1

[[2]]
[1] 2

[[3]]
[1] 3

[[4]]
[1] 4

[[5]]
[1] 5

> sessionInfo()
R Under development (unstable) (2017-01-14 r71969)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 16.04.1 LTS

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] stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
[1] BiocParallel_1.9.4   BiocInstaller_1.25.3

loaded via a namespace (and not attached):
[1] compiler_3.4.0 tools_3.4.0    parallel_3.4.0

ADD REPLY • link 7.3 years ago Martin Morgan 25k

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Entering edit mode

> library(BiocParallel)

> param = MulticoreParam()

> param

class: MulticoreParam

bpjobname:BPJOB; bpworkers:158; bptasks:0; bptimeout:Inf; bpRNGseed:; bpisup:FALSE

bplog:FALSE; bpthreshold:INFO; bplogdir:NA

bpstopOnError:FALSE; bpprogressbar:FALSE

bpresultdir:NA

cluster type: FORK

> bplapply(1:5, function(i) i, BPPARAM=param)

[[1]]

[1] 1

[[2]]

[1] 2

[[3]]

[1] 3

[[4]]

[1] 4

[[5]]

[1] 5

> sessionInfo()

R version 3.2.3 (2015-12-10)

Platform: x86_64-pc-linux-gnu (64-bit)

Running under: Red Hat Enterprise Linux Server release 6.7 (Santiago)

locale:

[1] LC_CTYPE=ja_JP.UTF-8 LC_NUMERIC=C

[3] LC_TIME=ja_JP.UTF-8 LC_COLLATE=ja_JP.UTF-8

[5] LC_MONETARY=ja_JP.UTF-8 LC_MESSAGES=ja_JP.UTF-8

[7] LC_PAPER=ja_JP.UTF-8 LC_NAME=C

[9] LC_ADDRESS=C LC_TELEPHONE=C

[11] LC_MEASUREMENT=ja_JP.UTF-8 LC_IDENTIFICATION=C

attached base packages:

[1] parallel stats graphics grDevices utils datasets methods

[8] base

other attached packages:

[1] BiocParallel_1.4.3

loaded via a namespace (and not attached):

[1] futile.logger_1.4.3 lambda.r_1.1.9 futile.options_1.0.0

>

ADD REPLY • link 7.3 years ago JunLVI ▴ 40

score 0 · Answer 1 · 2017-01-22

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Entering edit mode

Ryan C. Thompson ★ 7.9k

@ryan-c-thompson-5618

Last seen 8 months ago

Scripps Research, La Jolla, CA

Based on your comment, I think you're seeing the same problem that I sometimes see when using MulticoreParam, where it causes R to hang forever. (Bioc maintainers: I only see this sometimes and haven't figured out how to reproduce it yet.) My suggestion would be to try another parallel param, such as DoparParam():

library(parallel)
library(doParallel)
registerDoParallel(cores=detectCores())
library(BiocParallel)
register(DoparParam())

Then you should get parallel read counting.

ADD COMMENT • link 7.3 years ago Ryan C. Thompson ★ 7.9k

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Entering edit mode

Hi Ryan, Thanks for your reply. I retried MulticoreParam, somehow it worked. (why? I do not know). I will try DoparParam when MulticoreParam do not work. TKS!

ADD REPLY • link 7.3 years ago JunLVI ▴ 40