DD-PCR--Annealling temps

douglas l feinstein dlfeins at cumc.cornell.edu
Fri Feb 18 09:49:08 EST 1994


Concerning 10-mers and oligo dT primers for PCR: How is it that a random
10-mer with a calculated (at+gc) Tm of about 30oC can serve as primer
when the Tanneal is usually 37oC? Same goes for the oligo dT?
We are playing with dd-pcr, but rather than use different RNA as starting
material, we are using 2 different cDNA libraries, since we have already
made these (e.g. control and treated cells). Has anyone done this
already? In this way we only are using the 10-mers. In fact, from what
I've seen at some meetings, many of the products subseqeuntly sequenced
turn out to be primed only with the 10-mers, not with oligo dT

Bye the way, I keep having problems trying to post a follow up to
messages, and have to resort to posting new messages. ANy ideas?

thankx
dlf
.
Doug Feinstein																	 |  Voice:   212 570-2900
Dept Neurobiology															|	Fax:      212 988-3672
Cornell University Medical College    |  E-mail: dlfeins at cumc.cornell.edu
We're not the best at what we do, but we're the only ones who do it...



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