Question: Validation of RNASeq Data - How to validate RNASeq DEGs using qPCR
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gravatar for Sreeraj Thamban
2.7 years ago by
Indian Institute of Science Education and Research
Sreeraj Thamban150 wrote:

Hi, Biostars! Can anyone tell me how to validate the RNA-Seq Differential gene expression data using qPCR? How many genes should I select for qPCR validation? Is there any criteria for selecting the differentially expressed genes for qPCR validation? and finally what is the best method to visualize/represent the correlation between RNA-Seq and qPCR data?

Thank you all!!

rna-seq deseq2 • 2.3k views
ADD COMMENTlink modified 2.7 years ago by lessismore880 • written 2.7 years ago by Sreeraj Thamban150
1
gravatar for lessismore
2.7 years ago by
lessismore880
Mexico
lessismore880 wrote:

Hey! It's highly recommendable to validate at least 20 genes. I'd suggest to choose the ones interesting for the topic you are addressing in your study and ones completely random, or for which you observed an interesting behaviour. The method is a simple Pearson correlation between the two groups: 1: RNA-Seq data and 2: qPCR data. If you got a Pearson correlation value of min 0.7 consider your validation already quite good! Good luck!

ADD COMMENTlink written 2.7 years ago by lessismore880

Hi lessismore, Thank you for your answer. Is there any standard guideline for valiadting the RNASeq data using qPCR?

ADD REPLYlink written 2.7 years ago by Sreeraj Thamban150

The wise way is to have a look to the latest papers in your topic and to the related journal requirements. Then you can figure out what do you precisely need for your purpose

ADD REPLYlink written 2.7 years ago by lessismore880

Hello!, May you give to me some reference about these?

thanks!

ADD REPLYlink written 20 months ago by maperezm.medi0
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