ELECTROPORATION of E. coli

Brian Foley brianf at med.uvm.edu
Mon Jan 24 14:54:38 EST 1994


Seth Findley (findley at u.washington.edu) wrote:
: Pardon this probable FAQ, but...

: Would anybody have any tips for optimizing electroporation of E. coli?
: Is 1x10e10 efficiency commonly obtained?

	Not in my hands.  I have a hard time getting better than 1.0 x 10e9
transformants per microgram of pBluescript.  I use DH5alpha and XL1-Blue
E. coli strains.

: Does the growth medium make a difference?
:  
	I found the most difference in the O.D. of the culture, and keeping
it on ice throughout the washes.  A low O.D. and thus early log phase
growth was important.  Also cooling the cells on ice before spinning them out
of media and into the low ionic strength washes.

: We are using both DH5a and DH10b. 


: Thanks, in advance.

: Seth

--
********************************************************************
*  Brian Foley               *     If we knew what we were doing   *
*  Molecular Genetics Dept.  *     it wouldn't be called research  *
*  University of Vermont     *                                     *
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