competent cell preparation
mbzspd at granby.nott.ac.uk
Tue Oct 31 03:32:47 EST 1995
I think the method you are looking for was described originally in
Chung, C.T. and Miller, R.H., (1988) Nucl. Acids Res. 16, pp3580.
It's based on a PEG/DMSO/Mg2+ treatment and it works great. There have
been some more up to date versions (from the same authors I believe), one
of which was indeed in PNAS (PNAS (1989) 86, pp2172 - ??) and there is a
comprehensive report in 'Methods in Enzymology' volume 218 I think.
We use it a lot here as our standard method for getting competent
cells....I can routinely get ~5 x 10^7 cfu/ug pUC19. Not quite as high as
they claim but good enough for most things :)
If you want any more details, drop me a line via email.
Dr. Simon Dawson TEL:+44 (0)115 9249924 ex. 44789
Dept. of Biochemistry FAX:+44 (0)115 9422225
Queens Medical Centre Email:Simon.Dawson at .nott.ac.uk
Clifton Boulevard URL: http://www.nott.ac.uk/~mbxspd/Simon.html
U.K. "Back off man, I'm a scientist!" - Bill Murray.
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