DD RT-PCR

Joel Guss jg183 at columbia.edu
Tue Nov 25 21:15:48 EST 1997


Hello,

I was wondering if there is anyone here who works (successfully or
unsuccessfuly) with differential display RT-PCR, especially in brain
tissue and might be able to share their real-life experience with 
the technique.

Namely, how "hard" is it to get it to work?
How did you choose your primer pairs (how many and which)?
PCR conditions?
How difficult is it to visualize/extract results?
How much time does it usually take to get it to work properly?

All of the review articles I have read made it seem like a great
technique but pointed out so many problems that almost make us
hesitant to try it out.
Can anyone suggest any really good protocols or very recent reviews
critically comparing different techniques or even kits and the 
advantages and disadvantages of each?
Thank you very much for your time; any help is really appreciated!
Please reply by e-mail if possible, or I will try to check the
newgroups.

				Joel Guss
				Columbia University
				jg183 at columbia.edu



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