transducing recA w/o tetR ?

Hank Seifert 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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