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Gene expression

Gene expression - PCR - Polymerase Chain Reaction Forum

Gene expression - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 07-20-2012, 07:42 AM
Pipette Filler
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Default Gene expression



Dear all:
Currently I’m working on an experiment for evaluation of IL-17 (interlukin-17) expression, I used house-keeping gene (GAPDH) as an endogenous control. I started with RNA and used a single step RT-PCR mix and a realtime PCR. Now I’ve the Cts and not quite sure what to do with the analysis. I don’t have a standard curve for this gene, but I have another one. I’ve been advised to use relative quantitation (ΔCt). But I’m not familiar with the steps of calculating or how I’m going to consider this gene expressed or not. Also there’re some software tools for this matter does anyone here have one?
Thanks for your help

Last edited by Salome egypt; 07-20-2012 at 07:43 AM.
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Old 08-02-2012, 01:25 PM
Pipette Filler
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Default Re: Gene expression

Hi,
you can find out the method in this paper "A new mathematical model for relative quantification in real-time RT–PCR" by Michael W. Pfaffl
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