magic mp & sequenase

michael coyne michael.coyne at channel1.com
Thu Dec 3 01:21:30 EST 1992


Hello all...

In advance of some experiments to be performed by a co-worker (and at
the request of my boss), I would like to find out if anyone has tried
using plasmid DNA recovered with Promega's Magic MiniPrep kit directly
for use with the Sequenase kit.

The plasmid we're recovering is small (apx. 6 kb), yield and cleanliness
look good on agarose gels, but the Sequenase kit has been (in our hands)
somewhat fussy about the DNA used.

I remember, for example, a post here not too long ago discussing the use
of alkaline-lysis (Birnboin & Doly) mini-prep DNA with the Sequenase
kit, after cleaning it up with glassmilk.  This protocol, as I recall,
advised using two times the recommended volumes of the Sequenase kit
reagents when using this method.

Has anyone had similar experience tweaking the kit to accept plasmid DNA
recovered with the Magic MiniPrep kit?

Any information as to the viability of this approach, and suggestions
for its successful implementation, will be greatly appreciated.

Thanks...

Mike
michael.coyne at channel1.com
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Channel 1 (R)   Cambridge, MA



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