BioRad's Prep-A-Gene, try it, you'll like it

Bruce Roe broe at aardvark.ucs.uoknor.edu
Fri Apr 1 09:32:00 EST 1994


Hi,
	To all you folks out there who want a kit for preparing your favorite
dsDNA, I suggest you contact Bio-Rad and order their Prep-A-Gene kit.

We have successfully used Prep-A-Gene to isolate plasmid, cosmid and P1 DNAs.
The plamsids, when isolated using Prep-A-Gene and the Prep-A-Gene 24 well
filtering apparatus, serve as excellent templates for both radioactive and
fluorescent labeled primer, Taq cycle sequencing on the ABI sequencer.  We
scaled up the protocol for using Prep-A-Gene to purify DNA from 50 ml cultures
employing a centrifugation-based approach rather than a filtering-based
approach, and this dsDNA is an excellent template for obtaining sequence with
Sequenase and the ABI fluorescent terminators.  When isolating cosmids and P1s,
the centrifugation-based approach using Prep-A-Gene, the dsDNA is of extremely
high quality, has very little, if any host genomic DNA, is easily cut with
restriction enzymes, sub-cloned, etc., and is almost totally in a supercoiled
form.

The Prep-A-Gene resin is diatomaceous earth, and Bio-Rad is the patent holder
for the use of this material in DNA purifications.  Diatamaceous earth is
available from Sigma, and I have previously posted our protocols.  However,
Prep-A-Gene is specially treated and quality controlled for binding capacity,
lack of nucleases, etc and I strongly recommended it for those who don't want
to fool around with pre-treatment, making the necessary buffers, and still
having something that is not specifically sold for DNA purifications and
therefore not guaranteed to work as advertised.

Disclamer:
At one time I did consult for and collaborate with several scientists at
Bio-Rad, and therefore have some friends there, but at present have no
affiliation with Bio-Rad and nothing personal to gain from this posting.

Cheers......bruce
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