Is it possible in DiffBind to specify sizeFactors for DESeq2 analysis?
2
0
Entering edit mode
@nfursovamsu-11039
Last seen 8.4 years ago

Hi! I was wondering if there is a way to provide DiffBind with previously defined sizeFactors? I am using a calibrated ChIP-seq approach and can calculate normalisation factors (with DESeq2, for instance) using reads mapping to my reference genome and I would like to use these normalisation factors as sizeFactors when I perform differential analysis on my experimental genome.

diffbind normalization • 1.1k views
ADD COMMENT
0
Entering edit mode
Rory Stark ★ 5.2k
@rory-stark-5741
Last seen 5 weeks ago
Cambridge, UK

There is no easy way to do this from within DiffBind. While there are some tweaks that can be done to modify the library size used in the calculation, I think you want more fine-grained control than that. This is a good example of a case where you'll want to be using the underlying analysis package (DESeq2 or edgeR) explicitly. You can use DiffBind to calculate consensus peaks and do centering/counting, and export this to the differential analysis package using dba.peakset() with bRetrieve=TRUE. You'll want to set the read score appropriately, e.g. to DBA_SCORE_READS.

We are looking at possibly adding a set of features to enable more explicit control of normalization in a future release.

Cheers-

Rory

ADD COMMENT
0
Entering edit mode

Thank you very much, it was very helpful!

ADD REPLY
0
Entering edit mode
Rory Stark ★ 5.2k
@rory-stark-5741
Last seen 5 weeks ago
Cambridge, UK

The latest release of DiffBind includes a new interface function, dba.normalize(), that allows for much more flexible normalization. The updated vignette includes a detailed exploration of the impact of different normalization schemes.

ADD COMMENT

Login before adding your answer.

Traffic: 555 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6