Minipreps and Automated Sequencing
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!
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
Tennis Court Road, Answerphone & alt. FAX 333661
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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