Touchdown pcr

[Michael Kertesz]

Hi Phillippe,

Touchdown gives me much better specificity, especially when the two 
primers have Tm's that are way apart. I start at an annealing temp. about 
10-15 deg. too high, then come down with it one degree per cycle, before 
finally running a "normal" PCR for 15-25 cycles at the end. We have a 
Trio-block, though, where it is dead easy to program, though...

regards

Michael 


Philippe Lefebvre wrote:
> 
> Hi everybody!
> Does anyone has already used touchdown pcr, and could say more about pros and cons
> about this method.  May be is there even a protocol available out there?
> Thanks in advance for your help!
> Dr Phil Good

-- 

Michael Kertesz

ETH-Microbiology,
ETH-Zentrum/LFV,
CH-8092 Zurich, Switzerland

tel: +41-1-632 3357
fax: +41-1-632 1148
e-mail: kertesz at micro.biol.ethz.ch