DDRT-PCR

Paul C. Grimm MD grimm at cc.UManitoba.CA
Mon Dec 16 16:11:34 EST 1996


Dr Alok Adholaya wrote:
> 
> Dear Netters,
> 
> I would like to know if there are any ways of conforming the
> differentially expressed clones isolated using Differentail display
> PCR (DDRT-PCR) other than the standard Northern blot. I am using a
> system where the availability of total RNA is a premium ( Not to
> talk of Poly A RNA!!!). I am looking for ways and means to check
> for the differential expressed nature of the clones where i would
> not require large quantities of RNA.
Why don't you try the Virtual Northern Technique using the Capfinder Kit
technology from Clontech?


> P.S :  I want to know the opinion of those of you who are
> using/used  Silver staining  DNA sequencing system of Promega Corp
> ? Is it as sensitive and easy as made out to be ? Does it "really"
> replace  the radioactive mess which we exopse ourself when we do
> sequencing.

Yes
I use the recipe (with a few modifications) from Promega and it is just
fine for sequencing and for differential display. There are some
Biotechniques articles on using silver staining for DDRT-PCR
-- 
Paul C. Grimm MD
Assoc. Prof. Pediatrics & Immunology
University of Manitoba
Winnipeg Man, Canada
R3A 1S1

"Having a smoking section in a restaurant is like
 having a peeing section in a swimming pool!"




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