transducing recA w/o tetR ?
h-seifert at nwu.edu
Wed Jul 6 17:19:53 EST 1994
In article <2vekekINN35s at s-crim1.dl.ac.uk>, mbrgw at s-crim1.dl.ac.uk (R.G. Walters) says:
>[On fusaric acid selection...]
>>: Roughly 80-90% of colonies on these plates were (in my hands) Tet sensitive.
>One thing I found helped was incubating at 30C. The selection seems more
>effective if the cells are growing/dividing more slowly.
>Oh - and of course you have to grow the cells in the absence of Tet prior
>to plating them out. Else all the ones you want are dead :-(
>Robin Walters. Robert Hill Institute, Sheffield UK.
>A fact is an opinion that everyone agrees with.
Many years ago, I used the Bochner method. The trick was not to plate too many
cells on each plate. If your frequencies are better than 10^-6 then it worked well enough
to enrich the tet^s to about 50% of the colonies.
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