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RT-qPCR of selected DEG: which dilutions in calibration curve?

RT-qPCR RNA-seq rna-sequencing

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#1 Bernardo

Bernardo

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Posted 04 December 2013 - 10:09 AM

Hi all, this is my firs post:
 
I've seen in MIQE guidelines that calibration curves a needed when performing RT-qPCR. I'll use genomic DNA to perform these calibration curves.
 
My question is that I don't know the range of concentrations to test.
 
The genes I will check for differential expression have logFC between 2-8 (base 2 log).
 
50 ng/μL is the sample concentration I will have when performing the Comparative CT Experiment, but 98% is ribosomal RNA.
 
Please let me know your suggestions, possible reference books/papers to revise and if this has been already discussed.
 
Thanks for you help. Best regardssmile.png , Bernardo

Edited by Bernardo, 04 December 2013 - 10:57 AM.






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