RT-qPCR: Calculation of Pairwise variations to determine the optimal number of reference genes

Question

[mohammedtoufiq91]

Hi,

I am using geNorm, Normfinder, RefFinder, and BestKeeper to identify and select the appropriate reference genes for the normalization in the RT-qPCR analysis. I was trying to identify the tools to calculate the pairwise variations (Vn/n + 1) to determine the optimal number of reference genes to calculate the V value such as V2/3, V3/4, etc. Should this calculations needs to be performed manually, and how can this be performed or any tools could specifically perform this task. Please assist me with this.

Best Regards, Toufiq