Trivial transformation improvement

Frederik Börnke ricky_boernke at gmx.net
Fri Feb 11 01:59:25 EST 2000



Dima Klenchin schrieb:
> 
> :In article <38A25F8E.E25F66D1 at gmx.net>, Frederik Börnke
> :<ricky_boernke at gmx.net> writes
> :>Yeah, but after back to the ice you probably do a recovery step at 37
> :>deg,
> :>don't you?
> :>BTW I do my recovery step usually in a shaker not in a waterbath.
> :
> :The same applies to electroporation. What is the best temperature,
> :treatment and time for post-recovery? I've seen all sorts in the
> :literature from RT to 37C to shaking/non-shaking from 10  mins to 60
> :mins. I've also seen it said that with Amp one needs recovery but for
> :Kan one doesn't.
> 
> How is that possible? In either case, don't you need some "recovery"
> time to actually make enough protein that confers resistance?
> 
>         - Dima

I haven't tried that but it sounds plausible. Maybe the cells don't
grow right away after plating they actually should be quiete enfeebled
after they come from the solution were they have been made competent
with. BTW, I looked up the Novagen catalogue an they say for their
AccepTor Vector kit there's no incubation step necessarry for the
version confering amp resistence, haven't tried it though.

Ricky


*******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiology
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
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