Hi!
I am trying to plot DNA binding profiles of my ChIP-seq bw files using Deeptools plotProfile. I generated the matrix using the computeMatrix reference-point. I used some publicly available bed files as my regions of interest, but, some of them resulted in a strange pattern as you can see below.
I aligned my data to the mm10 reference genome with the default options of bowtie2. I generated and sorted the bam files with samtools and used Picard to remove duplicates. I then used the following commands from deeptools.
bamCoverage -b sample1.bam --effectiveGenomeSize 2494787188 --normalizeUsing RPKM -p max --extendReads 150 -o sample1.bw
computeMatrix reference-point -S sample1.bw IP.bw -R regions.bed --referencePoint center -a 3000 -b 3000 -out sample1.tab.gz --skipZeros -p max --samplesLabel sample1 IP -bs 100
I used the peak files from the following links:
https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE116601
https://www-ncbi-nlm-nih-gov.proxy1.lib.uwo.ca/geo/query/acc.cgi?acc=GSE155890
Using the computeMatrix scale-regions did not solve the problem too. What is the reason for this problem and how can I solve it?
Thanks!
1 answer
I am having the same issue dealing with PRO-seq data, and the heatmap doesn't support this to be a true profile. Wonder if you have solve the problem or not?
Log in to answer this question.
Can you link to the bed files you used? You may also want to add the commands you used to align your ChIP-seq reads, and also your deepTools commands.
Thanks for your reminder! I added the links and commands.
What are those BED files supposed to represent? ATAC-seq peaks?
No they are ChIP-seq peaks that might bind to the same regions as mine