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Old 11-02-2012, 10:06 AM   #1
Location: Texas

Join Date: Apr 2012
Posts: 48
Default how to calculate the mean sequencing depth of RNA-seq

Dear users

I tried to calculate the mean sequencing depth of RNA-seq. Firstly, I utilized GATK to calculate that with the whole genome as reference. It turned out that some samples with around 15 million mapped reads only have around 2.5* mean sequencing depth. That looks to me weird. Some people suggested me to calculate it by hand, such as: (number of mapped reads * read length)/total length of the species' transcriptome

It looks reasonable. How do you calculate the sequencing depth of RNA-seq?
I will appreciate your comments very much!
wangli is offline   Reply With Quote
Old 02-18-2014, 06:38 PM   #2

Join Date: Jan 2010
Posts: 52

You should calculate the sequencing depth of a gene or an exon. It doesn't make sense to have a number representing the overall sequencing depth of RNA-Seq. DNA and RNA-Seq are different.
woodydon is offline   Reply With Quote
Old 02-19-2014, 03:56 PM   #3
Location: Europe

Join Date: Nov 2011
Posts: 52

You could use e.g. Cufflinks or DEseq to calculate the library depths, at least that would provide you a scaling factor for inter-library comparisons and normalization. I think the "sequencing depth" as per DNA-seq is not comparable in this respect at all.
puggie is offline   Reply With Quote
Old 02-19-2014, 05:44 PM   #4
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Location: Pathum Thani, Thailand

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The most informative coverage metric for RNA-seq is the number of genes with x number of reads, e.g. 80% of the known genes had at least 10 reads mapping to them, 95% of the known genes had at least one read map.
Jeremy is offline   Reply With Quote

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