actin as internal standard in RT-PCR
jirving at aol.com
Sun Oct 8 03:48:51 EST 1995
<In semi-quantitative RT-PCR, is it valid to use beta-actin as an
internal standard when comparing the expression of a gene in DIFFERENT
TISSUES? Or is it only valid when comparing the expression of a gene in
a specific tissue at different developmental stages?
If the answer to question 1 is "no", what internal standard is best to
Don't use actin , since actin expression is cell-cycle dependent; it's
best to use GAPDH, although this also has some variability in rare cases.
I have used GAPDH for my external standard in quantitative RT-PCR for five
years or so and it works well for me with minimal variability. I wouldn't
recommend using it as an internal standard in the PCR reaction itself of
course, since this would give you much more variable kinetics. There have
been a number of papers in the past few years regarding this topic; let me
know if you need some references. Good luck!
More information about the Methods