Quick transformation protocol wanted

Troels Wind wind at biobase.dk
Thu Feb 13 11:41:16 EST 1997


I haven't heard of what you describe, but alternatively you can
try the protocol of Pope and Kent (Nucl Acid Res, vol. 24, 536-7, 1996).

In short:
Harvest cells from 50 ml, OD=0.7-0.8.
Chill on ice, resuspend in 25 ml ice-cold 0.1 M CaCl2.
Chill on ice 30 minutes, harvest and resuspend in 2.5 ml 0.1 M CaCl2,
include 10% glycerol if cells are to be stored at -80C.
Store as 100 ul aliquots at 4C or -80C.

I dont remember for how long the cells are good at 4C, its in the
original paper.

Transform by adding 1-10 ng DNA and incubate on ice for 5 minutes.

Plate on preheated plates.

I use this method when shuttling plasmids between my favorite
strains, its not as quick-and-dirty as what you asked for, but it's
the quickest I've heard of...

Troels Wind
Aarhus University
Denmark

A.P.N. de Boer (deboer at nefeli.imbb.forth.gr) wrote:
: Hi,
: Once upon a time, I learned a very quick (and dirty) transformation
: protocol. It could be used to transform strains with plasmid DNA (which
: doesn't need a high transformation efficiency). It was very quick and and
: just entailed mixing some bacteria (colony from plate or pellet or
: anything), DNA and the transformation mix and the mix could be plated.
: I can't find the protocol anymore (moved labs). Does anyone here know of
: such a protocol ?
: Thanks, Thon de Boer



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