Hello,
Im working with co-infection PRRS virus/Swine influenza virus, i'm waiting for RNA-seq data, so please I need to know Is it necessary to confirm the RNA-seq results by quantitative RT-PCR and westerblot?
Thank you so much
Dear Zeineb,
First, you don't need to confirm all your RNA-seq results by qPCR. If you have a good coverage and a robust bioinformatic pipeline, I would trust your results to establish your list of interesting genes.
From your question, I am guessing that you'll be looking at differential gene expression between your infected and uninfected groups?
First, I would establish a list of candidate genes based on the RNA seq results.
Then, I would make a replicate experiment (not the original samples that were sequenced) to validate your subset of candidate genes.
Finally, I would only reconfirm by qPCR and WB the genes that you will investigate further in subsequent functional assays.
Hope that helps!
Nicolas
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