Filter mating

Michael Benedik bchs1b at Elroy.UH.EDU
Thu Jul 9 22:37:05 EST 1992


In article <1992Jul9.122641.29814 at urz.unibas.ch>, herzer at urz.unibas.ch writes:
>
>I would like a protocol or reference for filter mating.  I am trying to 
>transfer (conjugatively) a plasmid construct from one E. coli strain (
>which contains "tra" genes) to a wild-type E. coli strain.  Any help
>would be greatly appreciated.
>                               Thanks,
>                                      Pete 
>                                      "Herzer at urz.unibas.ch"
>
>
>
For transferring between E.coli strains you usually do not need to do
filter matings, which are designed for very low rates of transfer. I do one
of the following:
(1) If reasonable transfer rates are expected, I just add 0.1ml of each
culture to the bottom of a sterile test tube and incubate at 30 or 37
for a few hours then plate on selective media
(2) for an easy facsimile of filter mating I just spot a drop of each
strain on top of each other on an LB plate and incubate for a few hours
to overnight. THen scoop up the cells and either streak directly on selective
plates or resuspend a large scoop in sterile saline and plate dilutions
as appropriate.


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



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