mini-preps: NOT kits.
David L. Haviland, Ph.D.
HAVILAND at KIDS.WUSTL.EDU
Tue Oct 10 12:32:29 EST 1995
In <gustilo-0710952201520001 at ts11-14.upenn.edu> gustilo at pobox.upenn.edu writes:
> ABI has a user bulletin on how to go about doing a plasmid prep for their
> automated sequencer. I believe it is User Bulletin #8.
> It is essentially an alkaline lysis prep as described in Promega's User
> Manual with the addition of a 13% PEG 8000 precipitation. They also
> recommend using Terrific Broth in place of LB to grow your overnights to
> increase the DNA yield using the same volumes.
Agreed! This protocol also works well. However, I'd like to render a
personal observation and I'd be interested in any feedback.
*In my hands*, I've found that PEG solutions, autoclaved or not, are only
good for 6 or so months, then they "fall" off. By that I mean that I
obtain no DNA from the PEG midi-prep. As far as I can tell it is an all or
nothing event - either I get DNA or I don't, so I've cultivated the habit
of making the PEG/MnCl solution every 3-4 months and the protocols work
well for me. Hope this helps.
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