Trivial transformation improvement

CSC un691cs at genius.embnet.dkfz-heidelberg.de
Wed Feb 23 06:57:44 EST 2000


On 23 Feb 2000, Moez Torki wrote:

> I never do the 37 C recovery step. Indeed bacteria grow slowly (about 20
> h), but you get the same yield as for the 37 incubation. Try it.

Uuuuhhhh, dangerous !
This has been on this newsgroup a 100 times. It depends on 
1. the strain you are using
2. what you are trying to clone in which vector
3. how many colonies you want: if one will do, o.k. ! But if 0 is too
little: hmmm

With several strains/experiments I have noticed a three fold rise in
transformants after a 1-2 hour incubation at 37C before plating.
Sometimes it was 3 colonies vs. zilch ! 
Makes sense too: the little buggers need a little time before they have
produced enough beta-lactamase. For those of you who don't want to think:
always include the 37C step.

Kind regards, 

Clemens Suter-Crazzolara, PhD
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