Trivial transformation improvement

ChenHA hzhen at freeuk.com
Thu Feb 10 19:43:28 EST 2000


"Dr. Duncan Clark" wrote:

> I've also seen it said that with Amp one needs recovery but for
> Kan one doesn't.
>

It's the reverse.  You need recovery time for Kan because it inhibit
protein synthesis, therefore without a recovery period the gene product
that produce Kan resistance is not made.  For Amp type antibiotics, the
cell wall synthesis is affected but cell only get lysed when the cell is
expanding or dividing.  There is therefore time for the cells to produce
beta-lactamase before the antibiotics cause any serious damage.  I
sometimes plate the cells straight after heat shock (if I'm short of time)
and it seems to work well.  However I have not made any comparison of
transformation efficiency with and without incubation.

>
> Duncan
> --
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
>
> Duncan Clark
> DNAmp Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
> http://www.dnamp.com
> http://www.genesys.demon.co.uk





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