kit for differential display PCR

Shahram Mori smori at nmsu.edu
Wed Nov 22 21:14:37 EST 1995


William Sun (william at neuro.usc.edu) wrote:
: Hello,

: I am interested in using differential display PCR as a technique to see
: differential RNA expression in brain tissue.  I need some recommendations
: for a kit that is reliable and not too expensive.  Please reply if you
: have any suggestions.  Thanks.

There are no kits, because 90% of the time this technique doesn't work....
I am fed up with it. I worked on it and my hair is all white.

I think it's primarily LUCK. yES flame me if you want, but I can point to 
100 people who this technique hasn't worked for for everyone that you 
point it worked for..... Complaining aside.

I use Perkin Elmer's GeneAmp kit. Basically I use the ragents and don't 
follow any of the protocol. I use Superscrip II (GIBCO) for RT to get a 
more efficient reaction. I do everything in triplicates and let my Rt 
reacion go for 2 hrs.
My problem is that I get beautiful gels and can reamlify Beautifully BUT 
when I do northerns-> Nada. Zip. Zilch.... They all show up positive in 
the control and the test condition...
This is where I think the LUCK comes from.. You have to do enough to get 
one or two to work for you. You may get lucky and get one the first time 
you do it or have horrible luck and do 50 and not get one...

Hey Paul Why not write about this in Trends in Biological Sciences....
Tell'em shahram is suicidal.. :)

Cheers
--
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Shahram Mori
Program in Molecular Biology
Department of Chemistry and Biochemistry Box 3C
New Mexico State University
Las Cruces NM
88003
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