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Boiling Method for Minipreps

Michael Benedik bchs1b at Elroy.UH.EDU
Thu Aug 5 00:06:52 EST 1993


In article <1993Aug4.221947.9602 at alw.nih.gov>, Jim Owens <jow at helix.nih.gov> writes:
>In article <9308032222.AA20703 at calshp.cals.wisc.edu> ,
>jchen at CALSHP.CALS.WISC.EDU writes:
>>As a matter of fact, I prefer HB101 as a
>>host because cells are easy to resuspend into the lysozyme buffer.
>
>Somewhere in my collection of references there is a report about lysozyme
>not being necessary, and even reducing plasmid yield, in some strains of
>_E. coli_.  So I could not help wondering if anyone has tried a boiling
>miniprep without adding lysozyme.
>
>I personally use the alkaline lysis method modified somewhat, but without
>lysozyme, for mini-, midi- and maxipreps of plasmids.
>
>Good luck,
>
>Jim Owens


As a matter of fact I just did that experiment today because we were
also discussing whether or not it was important.
Our result was significnalty higher DNA yield when lysozyme was added.
this was using strain TG1 and high copy plasmid pBluescript.
I strongly suspect the answer is strain dependent, but for TG1, which is
a very hardy strain, lysozyme really helped.


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 Michael Benedik				INTERNET: Benedik at uh.edu
 Dept. of Biochemical & Biophysical Sciences	
 University of Houston				BITNET: Benedik at uhou
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