TRANSFER OF PLASMIDS INTO BACTERIA
Ian A. York
york at mbcrr.dfci.harvard.edu
Fri Mar 3 16:43:55 EST 1995
In article <leach-0303951105040001 at med-pharm16.bu.edu> leach at bu.edu (Martin D. Leach) writes:
>Obtain some sub cloning efficiency DH5alpha cells (e.g. Gibco BRL)
>[ . . . ]
>then transfer to 42C for 45sec then replace on ice for 2mins
Or, if you read the instructions, try 37oC . . .
If you're really paranoid, order BRL's (EXPENSIVE) library or
(even more expensive) maximum efficiency competent E coli. Other
companies e.g. Invitrogen also sell competent bugs (just to show I have
no affiliation with anyone). -And go with the manufacturer's directions
for each variety.
Otherwise I agree completely with Martin here.
Ian York (york at mbcrr.harvard.edu)
Dana-Farber Cancer Institute, 44 Binney St., Boston MA 02115
Phone (617)-632-4328 Fax (617)-632-2627
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