protocol colony PCR?

Gerd  gerdn at ibg.uit.no
Fri Sep 10 07:14:45 EST 1999


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In article <7r7u75$m51$1 at info.service.rug.nl>, "Rene Haanstra"
<r.haanstra<nospam>@biol.rug.nl> wrote:


>Dear Netters,
>
>Does anyone have a protocol for colony PCR that works for most
>bacteria? I experience a lot of problems when i try to do a 16S PCR
>on colonies of bacteria. Two out of five colonies don't produce a product....
>Rene Haanstra
>
...
>

Would that be five colonies from the same strain? 
In that case: what's the difference between the colonies? Age? Size?...?
In my hands colony PCR "always" works, - if the colonies are fresh.  - Old
plates: just forget it!

If it's two of five different strains that do work, then the differences in
physiology also can contribute to the problem. Try testing different initial
denaturing temps; 95 C - 97 C for 5-8 min are what I have played with. Then
use the program you have developed. Good luck.

Gerd



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