Minipreps and Automated Sequencing

Christopher Walentas cdw1000 at bioc.cam.ac.uk
Fri Feb 3 20:05:06 EST 1995


Does anyone out there know what the best way to prepare low-copy number plasmid
(mini-prep scale) for ABI automated sequencing?  We've tried Promega Wizards
(Magic uesd to work but that's another story), but they don't like to yield
any sequence even with additional precipitations/ washed to remove excess salt.
We tried QIAGEN 20 tips-- they work great down the hall where they use pUC
and cosmids, but the yields for pKK223-3 and pGP1-2 derived vectors are
deplorable.  Folk here have even put CsCl banded DNA down a 20 tip to find 
that they are only able to recover 10% or so.  What we've settled on is either doing larger minpreps (or rather a bunch of 'em), or chloramphenicol.  Is there a better way?

As an aside, if Promega was in patent conflict with their Magic resin, who 
DOES have the patent so that I can go back to what used to work.

Please forward any and all suggestion to the email address below.

Cheers in advance!


Tripp

- 
Christopher D. Walentas

Centre for Molecular Recognition,		Tel.:
Sir William Hardy Bldg.,			within the U.K.  (0223) 333662
Department of Biochemistry,			outside       +44 (223) 333662
University of Cambridge,			FAX		        333345
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Cambridge, England  CB2 1QW

e-mail: c.d.walentas at bioc.cam.ac.uk

under the supervision of Professor R. N. Perham



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