HELP!! transformation problems
Frederik Boernke
boernke at nospam.ipk-gatersleben.de
Wed Aug 5 17:16:21 EST 1998
Frank O. Fackelmayer wrote:
>
> For routine cloning, where maximum competence is not an issue, we always use
> the TSS method of Chung et al.
> It is very reliable once you got it to work.
>
> PS: Do not try to freeze the bacteria. They will go down in competence at
> least a factor of 100. Well, that might still be ok for transformation of
> supercoiled plasmids, but does not work for ligations any more.
>
Hi!
When I get it right you always have to prepare fresh cells when planning
a transformation, haven't you? So, what is the advantage compared to a
protocol where you can freeze/store the competent cells for ages? In
this
case you prepare once a huge amount of bugs and from time to time you
pull some out of the freezer.
Gruss
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
http://www.ipk-gatersleben.de
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