Introduction of restriction sites via extended primers?

Peter Frank peter_frankde at
Fri Aug 13 16:54:13 EST 2004

I wrote:

>... For the primer-template hybrids the overhang will be irrelevant
>and not affect the annealing temperature.

I have to correct myself or at least be more specific. It will not be
relevant for the very first primer-template hybrids with the template
being the molecules one introduced into the PCR mix.
But for newly synthesized amplicons serving as template it will affect
the annealing temperature.


More information about the Methods mailing list