Entering edit mode
Anitha Sundararajan
▴
140
@anitha-sundararajan-6152
Last seen 10.2 years ago
Hi
I have been trying to use DiffBind to analyze our Chip-seq data and
have
been running into some errors repeatedly.
I first created a samplesheet.csv describing my samples and it looks
like this:
SampleID,Tissue,Factor,Condition,Replicate,bamReads,bamControl,Peaks,P
eakCaller
meio.1,meiocytes,H3K4me3,N,1,M_meiocytes_H3K4me3.bam,InM_input_meiocyt
es.bam,meio.vs.in.rep1.def_peaks.bed,MACS
seed.1,seedlings,H3K4me3,N,1,S_seedling_H3K4me3.bam,InS_input_seedling
.bam,seed.vs.in.rep1.def_peaks.bed,MACS
I only have two samples (and their respective inputs) with one rep
each
and the peaks were called using MACS v2. The peak caller generated
.bed
files which was used in DiffBind.
I defined the working directory in R first.
I then read the sample sheet in :
> H3K4.B73=dba(sampleSheet='samplesheet2.csv',peakFormat='bed')
>H3K4.B73
2 Samples, 38870 sites in matrix (45304 total):
ID Tissue Factor Condition Replicate Peak.caller Intervals
1 meio.1 meiocytes H3K4me3 N 1 MACS 44124
2 seed.1 seedlings H3K4me3 N 1 MACS 41596
generated a plot,
> plot(H3K4.B73)
And then when I tried to perform dba.counts, it continuously fails on
me. I went through the thread to find similar posts and could not
find
a solution. I tried the floowing command:
> H3K4.B73=dba.count(H3K4.B73, minOverlap=3)
and this,
> H3K4.B73=dba.count(H3K4.B73, minOverlap=3, bLowMem=TRUE)
> H3K4.B73=dba.count(H3K4.B73, minOverlap=3, bLowMem=FALSE)
And they all failed.
Please let me know if you have any insights on it.
Thanks so much for your help in advance.
Anitha Sundararajan Ph.D.
Research Scientist
National Center for Genome Resources
Santa Fe, NM 87505