On again off again PCR

[Andrew Atwell]

Tara Paton wrote:
> 
> Hi,
> 
> I'm doing PCR of selected pieces of an avian mitochondrial
> genome and am having trouble with some of the amplifications since the
> primers I'm using are not the best match for my template.  Attempts at
> amplifying these fragments have failed miserably although some have
> worked in the past (ie they are not imposible). Are there any
> suggestions of strategies I can use to improve the frequency of
> success?  I'm currently using the profile:  94 (45 sec)- 50 (45 sec) -
> 72 (1.5 min) and the fragment lengths are no longer than 1.2 kb.  These
> amps have worked in the past (barely) but I need a better strategy than
> pure luck!
> 
> Thanks for your time,
> Tara Paton
SCIENCE!  Got to love good old Mother Nature.  If it always worked they
wouldn't call it RE-search.  Good luck try some fairy dust.
Andy
-- 
Andrew Atwell			"Yes officer, I am aware that I was
Palo Alto, CA 			traveling at 120 m.p.h"  -Jetta VR6
Andrew_Atwell at hp.com