Hello everyone,
So I have ChIP seq data regarding to the two key TFs in mESC namely Oct4 and Sox2. For quantitative purposes, exogenous ChIP seq data (Dm3) is also added and I want to carry out the normalization using ChIPSeqSpike package. However, subtracting the experiment and the input data using inputSubtraction function returns to error. It consistently said that the chromosomes differ between my experiment and input data. The code and error are as follow:
cs <- inputSubtraction(cs)
Reading the input file
Subtracting input to experiment
Processing Oct4_KO
Reading bigWig file
Error in FUN(X[[i]], ...) :
Chromosomes differ between input and experiment
In addition, in the traceback it is stated that the error might occur due to difference procedure in generating the bigwig files. Thing is, all the bigwig files are generated using the exactly same way from bedgraph. the example codes are as follow.
nice -n 5 genomeCoverageBed -bg -ibam ~/15912_mm9.q1.rmM.rmdup.s.bam -g ~/mm9.sizes > ~/mm9_KO3_bg_Oct4.bedgraph
nice -n 5 genomeCoverageBed -bg -ibam ~/15914_mm9.q1.rmM.rmdup.s.bam -g ~/mm9.sizes > ~/mm9_KO3_INPUT.bedgraph
for i in "mm9_KO3_bg_Oct4" "mm9_KO3_INPUT" ; do
echo ${i};
nice -n 5 bedGraphToBigWig ~/${i}.bedgraph ~/mm9.noHeader.sizes ~/${i}.bw;
done;
So can anyone please pinpoint where did I do wrong or how do I correct this error. all input is appreciated. thank you
Are your files in fixed steps?
Hello Nicolas,
I am quite new with this, could you please elaborate what do you mean by fixed steps?
In order to perform the input subtraction you need that your wig files are in fixed steps format, check this page: https://genome.ucsc.edu/goldenPath/help/wiggle.html
no I don't think so. okay I will try to fix the data first
so I tried to generate the fixedstep bigwig file using
then
but it still reproduce the error. May I know if the code is wrong or how can I properly generate fixedstep bw files?
The way I do it is to use bigWigToWig from UCSC and then you can try this R script:
Thanks Nicolas, I will try it first
Hi Nicolas, anyway for
can I use the chrome sizes file for that? because when I run it with my chrom sizes file as input, the chrom_length return to NA
what organism are you working with?
Hello, sorry for the late reply since I also working on another datasets
I am working with mouse genome and use mm9 to align my sequences data. so should it be the chromosome length of the mm9?
Keep the two first columns of the chromInfo table from ucsc: https://genome.ucsc.edu/cgi-bin/hgTables?hgsid=1020948187_TTkypQ5F8q84nAyf551tPR3wiXs9