CsCl preps for ABI sequencing reactions

Eric C. Anderson anderson at pharmdec.wustl.edu
Mon Mar 20 12:52:01 EST 1995


i was wondering if anyone had any information on using CsCl purified
plasmids (both straight and desalted) for sequencing using the Taq
terminator chemistry on an ABI autosequencer.

personally nobody in my lab uses this, we use Qiagen and Promega preps
(depending on size of prep and who is doing it), but we also ran a
departmental core facility and there are two labs who use CsCl purified
plasmids, usually desalted and whose reactions rarely work (usually about
40-50% of them work), whereas people in my lab, and others who use other
mini-prep methods get much better results.

since i'm not prepared to spend time with a cesium prep just to test a
hypothesis that doesn't affect me, i would like to know if anyone has had
similar problems or if these particular people are just doing something
wrong.  i have asked one of the people to do Qiagen preps of her strains
and repeat the samples just to compare, but if there is any other
information that people have that i can give to these labs (to save them
time, money and frustration) i would certainly appreciate it.

thanks in advance

eric

-- 
eric c. anderson                                
anderson at pharmdec.wustl.edu
dept. of molecular bio. and pharm.               (314)362-3963 (lab)
washington univ. school of medicine              (314)862-2435 (home)
660 s. euclid box 8103                           (314)362-7058 (FAX)
st. louis, mo 63110



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