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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 you suggestions and possible reference books/papers to revise.

Thanks for you help. Best regards, Bernardo

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I am not sure what you're trying to do. I think you want to measure gene expression level (amount of transcript), but then why would you use DNA as your reference? What do you want to calibrate with the DNA? –  yotiao Dec 5 '13 at 12:07

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