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how to normalize cDNA in semi-Q-PCR?

how to normalize cDNA in semi-Q-PCR? - Protocols and Methods Forum

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Old 08-13-2003, 01:08 AM
tanglin@auburn.edu
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Default how to normalize cDNA in semi-Q-PCR?




I am trying to use semi-Q-PCR to study gene expression in different tissues. but I still can not figure out ,how to
use internal control (beta-acin) to verify the equal amounts of RNA BE USED from different
tissues? or, another word, how to normalize my cDNA?
I got two procedures from "biologicalprocedures", and read again and again,
still confused, how to normalize cDNA from different sample RNA?
If you have experience, please give me some advice, I am highly appreciated.

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Old 08-27-2003, 06:52 AM
J. Hendriks
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Default how to normalize cDNA in semi-Q-PCR?

Hi,

I am using the LightCycler for several years now. The way we normalise is by
using beta-2 microglobulin (according to the literature this is a better
control)
I use the Ct crossing point of the two seperate reactions . Each reaction is
correlated to a standard and after doing this the two reactions of each
sample are correlated to eachother. This doesn't make sense reading it, but
I hope you will understand. If not let me know, I will send you an excell
template for your calculations.

Brenda
<[Only registered users see links. ]> wrote:
news:[Only registered users see links. ]...
tissues. but I still can not figure out ,how to
USED from different
again,
appreciated.
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