Diffbind GLM Design
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@liruiradiant-8906
Last seen 2.2 years ago
United States

Hello Rory,

Thanks for the Diffbind package, it makes DB analysis very easy.

However, I have the following question: How does DiffBind handle Input samples and how does it compare condition B with condition A? For example, I have a experiment design like this:

Condition    A    B
IP           2     2
IN           2     2

The number 2 indicates 2 biological replicates for each cell.

When DiffBind calls DESeq2, what contrast does it use internally to calculate LFC and FDR between condition A and B?

1. A_IP  - B_IP
2. (A_IP - A_IN) - (B_IP - B_IN) 
3. sth. else?

Thank you!

Ray

diffbind • 497 views
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I think when Input was not specified in the meta data, then '1' is used. When Input is provided, '2' is used?

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Rory Stark ★ 5.1k
@rory-stark-5741
Last seen 9 weeks ago
Cambridge, UK

The Input reads are not used in the DESeq2 modelling itself. By default, these reads are subtracted from the ChIP reads prior to modelling, you can control this using the bSubControl parameter when calling dba.analyze().

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