LB vs. TB?
Chia Jin Ngee
mcblab47 at leonis.nus.sg
Tue Nov 16 22:01:45 EST 1993
Hi to all!
In my institute, we have an in-house media prep room which stocks only
LB. So to get TB we have to make it ourselves, something we have not got
used to (heck! there are people making it for us why bother). So the
general procedure we use here to get more plasmid is to amplify them by
addition of chloramphenicol. Helps reduce the general bacterial bulk too.
As for TB, I've tried it and I can say I'm impressed :-) however, the
bacterial bulk in the end is huge! Trying to get good plasmid means you
will have to increase the reagent vols. So, I tried amplifying plasmids
grown in TB and the effect is great. The bugs loved the media that they
grew much more rapidly compared with LB. So by adding chloramphenicol even
at >0.6 A600/ml gives fantastic plasmid yield.
Lessons learnt? Always amplify plasmids for big cultures. Troublesome but
good results.
Blessings and regards
Genie
--
Jin Ngee, Chia Chemical Carcinogenesis Laboratory
(Genie, the OligoMan) Institute of Molecular and Cell Biology
mcblab47 at leonis.nus.sg National University of Singapore
Tel: (065)772-3797 Kent Ridge Crescent
Fax: (065)779-1117 Singapore 0511
"I'm Mr Chia not Ms Chia and I know Genie is a girl's name"-Jin Ngee, Chia
More information about the Methods
mailing list