mini prep procedure

the End jgraham at bronze.ucs.indiana.edu
Thu Apr 8 16:45:42 EST 1993


Kevin,

I've seen it consistently with both alkali and boiling methods. If you grow 
an overnight culture, and leave it in the fridge for another day and evening 
your miniprep will be real weak.

I imagine that if you let it go longer you will not see anything. What 
you should do is spin down your cultures and resuspend in your lysis buffer,
and then freeze them. Then they will be fine for quite some time.

Specualtions ? Well, sitting at stationary phase tends to alter the membrane
lipid composition of several bugs, apparently they are "toughening up"for
the duration. I guess that you either alter the ability to separate
plasmids for the membrane of get poor cell lysis.

You'll have to ask someone else how these cultures fare with the Supercharge
Extra-kleen miniprep columns :).

Jim



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