competent cell preparation

Simon Dawson mbzspd at granby.nott.ac.uk
Tue Oct 31 03:32:47 EST 1995


Hi,
   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.

                Simon.

-- 

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
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Nottingham
U.K.                       "Back off man, I'm a scientist!" - Bill Murray.



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