Quantifying differential expression

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Fri Oct 20 03:57:41 EST 2000

Robyn Roth wrote:


"I have a mutated gene on a plasmid, with the wild-type unmutated gene
still active on the chromosome.  Does anyone know of any methods where I
can determine the levels of expression of the 2 different genes?  The
mutated gene contains a single amino acid change, with a unique
restriction enzyme site engineered into it."

quantitative RT-PCR (see numerous posts about LightCycler etc) would be
the perfect thing for you.

But semiquantitative would also be an option, and you can distinguish
the products nicely with your inbuilt RE site.

I've never done it, but you should find a lot of literature and some
previous posts on this NG.

good luck


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