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how to compare mRNA levels of 2 different genes using real time?

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#1 krystle



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Posted 13 July 2010 - 07:32 AM

Hi, i wish to know the relative levels of 3 different genes in a biological samples.

I typically do the relative quantitation real time pcr where i obtain the relative fold changes for a particular gene of interest. I start off by doing rt-pcr to get cDNA. The cDNA is then used for real time pcr. However, this only allow me to look at differences in mRNA level of a particular gene in different samples.

How do i perform real time pcr to compare the mRNA level of different genes in one sample? I know i will need to do up standard curve but i can't seem to get the details on how to go about preparing the standard curve. What samples to use

Can anyone give me some information or any protocol so that i'll be able to perform the experiment?


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