Question: NGS plot error
gravatar for Constantine
5.3 years ago by
Constantine280 wrote:


I'm trying to plot the coverage of a bam file from a ChIP-seq experiment via NGSplot and I get the following error:

Configuring variables...Error in CheckRegionAllowed(reg2plot, default.tbl) :
  Unknown region specified. Must be one of: bed
Execution halted

This is the command I'm using:

ngs.plot.r -G mm10 -R tss -C myfile.bam -O mybamfile.ngs 

If I change the -R from "tss" to "bed" and specify a bed file I get from the PeakCaller software, it works just fine. However, I want it to take into consideration only the bam file

Any ideas why I'm getting this error message?



sequencing chip-seq next-gen • 3.4k views
ADD COMMENTlink modified 5.3 years ago by Fidel2.0k • written 5.3 years ago by Constantine280
gravatar for Fidel
5.3 years ago by
Fidel2.0k wrote:

The problem could be that your region's file does not end in .bed?

Otherwise, I suggest you to use deepTools instead (note: I am part of the developing team). Simply:

pip install deepTools

bamCoverage -b myfile.bam -o

computeMatrix -R tss -S -o matrix

profile -m matrix -o profile.pdf


Also, once you have a matrix you can plot a heatmap and add some clustring:

heatmapper -m matrix --kmeans 2 -o heatmap.png

You can take a look at the documentation ( for more information.

ADD COMMENTlink modified 5.3 years ago • written 5.3 years ago by Fidel2.0k

I'm mainly using deeptools (prefer it better) ;)

Just wanted to also use NGS to see whether I'm getting the same output...Guess I'll have to stick with deeptools then

ADD REPLYlink written 5.3 years ago by Constantine280

"computeMatrix: error: argument : invalid choice: 'tss' (choose from 'scale-regions', 'reference-point')" now this doesn't work .

But when i using the something like this

 "computeMatrix scale-regions -S -R gencode.v21.annotation.bed --beforeRegionStartLength 3000  --regionBodyLength 5000  --afterRegionStartLength 3000 --skipZeros -o matrix.mat.gz

It takes a lot of time but when i choose like chr21 this for my bed region it generates the matrix to reduce the matrix generation time ?

ADD REPLYlink written 21 months ago by krushnach80870
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