TOUCHDOWN: References needed

Beth Wapelhorst weazel at seanet.com
Wed Feb 8 23:20:08 EST 1995


A428ENDE at VM1.SARA.NL writes:
> 
>      I'm going to use this touchdown technique on PCR but because a silly prob
>lem, I can not get anything from our library. Will any of you please be so kind
> to send the best references about this? I don't mind to get them by news or e-
>mail, but probably it would be better by the news group so more people can get
>it.

I don't have the exact reference handy, but it's in Biotechniques. Did 
you just need a reference, or did you want to know how to do it? Here's 
how I do it: I get the best results from primers about 60C melting 
temperature.

5' 94C
Touchdown:
1' 94C
1' annealing - every cycle drop this one degree from 65C to 55C
1' 72C
Then like 30 cycles regularly: 1' 94C, 1' 55C, 1' 72C
7' 72C extension

This works perfect every time, with all our primer pairs. Every time 
someone mentions "hot start" I want to die laughing. Touchdown is the way 
to go.

sugarbeth
weazel at seanet.com



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