non specific pcr primiing

douglas l feinstein dlfeins at cumc.cornell.edu
Mon Feb 28 10:28:28 EST 1994


I have a pcr setup as follows:
						
						template:                          --------------------------
	         forward primer 1:             ----->
forward primer 2:                --->

Revererse primer:	                                                       
  <-----


As you can see, the primer1 and the reverse should work fine, while
foward primer 2 should not bind at all to the template. So... what is the
possibility that if I do the pcr with primer 2 and reverse primer I will
get a significant amount of product? What if the last 2 3' bases of
primer 2 overlapped with the beginning of thetemplate? I do this pcr at
62oC annealling, which is Tm-4oC of the primer 2.

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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