Quantitative PCR

jas allen jblue89 at hotmail.com
Mon Feb 20 14:18:39 EST 2006


Hi everyone out there...

I am going to attempt a quantitative (real-time) PCR for the first time
and am really confused about what to use to make the standard curve
with.  I want to quantitatively determine the differences in a gene
from human and mouse.  I just want to be able to say for example, gene
Q is expressed 5 times as much in these human cells than in these mouse
cells.

Can I create one standard curve which I can make these comparisons one
(for example, load a plate with samples from human, mouse, and some
kind of standard)  or do I need to create two standard curves and
separately quantitate these values?  How do I choose what to use as my
standard?

I am so confused.......any real-time PCR experts out there?  Any advice
would be great!

Thanks
Jas



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