PBR322 - maximizing yields

Michael Benedik bchs1b at Elroy.UH.EDU
Thu Aug 27 10:48:58 EST 1992


In article <wpcnbyr at lynx.unm.edu>, bhjelle at carina.unm.edu () writes:
>The postings about how to get great yields of pUC are all fine
>and good, but in most cases we get enough DNA from a pUC
>miniprep to keep us happy for a long time. 
>
>Unfortunately, some things can only be done in pBR-based
>vectors and I have been plagued with totally miserable
>yields of one particular example of same for more than
>a month now.
>
>I have tried chloramphenicol amplification (which I used
>to use in the old days when pBR was everywhere, but have
>lost my old recipe). Unfortunately, the major cloning
>manuals do not actually give understandable recipes for
>chloramphenicol amplification. The attempts I made to
>improvise did not seem to help.
>
>What are the best tricks for increasing yields of pBR-based
>vectors?
>
>Brian
>

the general recipe for chloramphenicol amplification is to add chloramphenicol
at a concentration of 50-100 microgram/ml. The precise concentration is
not that important. It is usually added when the culture reaches
a density of between 0.5-1.0 OD (A-600). Allow amplification to continue
for 12-18 hours. 

	---------------------------------------------------
	 Michael Benedik
	 Department of Biochemical and Biophysical Sciences
	 University of Houston
	 INTERNET: Benedik at UH.EDU	BITNET: Benedik at UHOU



More information about the Methods mailing list