E. coli cell free lysate

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Wed Apr 25 03:04:29 EST 2001


wolfsc at ibms.sinica.edu.tw ("Wolfgang Schechinger") wrote:
:Hi all, 
:
:I'd like to prepare a cell free lysate from E. coli for a 
:qualitative in vitro DNA replication experiment.  Is there some 
:easy procedure to try out (preferably lysing with triton 
:and some chelators like EDTA or something alike)?


Wolfgang, 

You are right (Re: detergents and chelators almost never
hurt), but if the issue is to be sure you are are really
"cell free", then you can never go wrong with a good ole'
*really* hard spin... (after a good lysis, naturally).

        - Dima




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