How many bases overlap in recombinant PCR?

Barry Mook barry at
Tue Oct 8 17:03:11 EST 1996

> I have used 9bp in each direction with success. 5bp maybe getting kinda short.
> Assuming 50%GC, a 5bp overlap would have Tm ~30C. Its worth a try, but you may
> have to play with your annealling conditions a bit (Temp, Mg++, etc.)
Good luck.
> Brett Lindenbach
Hi Brett

   Thank you for responding.  I'm considering ordering new primers and
trying again...however...I'd like to be sure that I'm not barking up ther
wrong tree.  When you did the 9bp overlap, how long was your entire primer
(approximately, of course).  I'll try annealing at lower
temperatures...thanks again.

Barry Mook
barry at

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