Hot Start PCR
weazel at seanet.com
Wed Sep 20 20:14:09 EST 1995
In my opinion, Hot Start PCR is a total waste of time. I use a protocol
called "touchdown PCR" which increases specificity without having to stop
and add stuff. (When you're doing 320 reactions a day, this is a big deal)
You just start off with an annealing temp that's about ten degrees
higher, and then decrease 1 degree every cycle to your optimum temp.
Finish the rest of the cycles at that temp. I choose all my primers to
have an annealing temp of about 60 degrees, and use the same touchdown
program every time.
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