Results: CsCl preps for ABI sequencing

Eric C. Anderson anderson at pharmdec.wustl.edu
Fri Mar 24 14:55:28 EST 1995


earlier this week i asked if anyone had any information about the use of
CsCl preps for automated sequencing using the ABI 373 and Taq terminator
chemistry.

i got a couple of replies that essentially said that they weren't very
useful for sequencing in this manner.  i checked with our ABI field rep who
said that due to the high level of incorporation of ethidium bromide, the
flourescently labelled ddNTPs won't incorporate very well into the
extension products.

as a test of this, i had one of the people who had been having trouble with
sequencing CsCl preps repeat the template/primer combinations using Promega
Wizard minipreps rather than CsCl preps.  they worked amazingly well when
compared to identical reactions with cesium prepped DNA.

just wanted to offer this bit of "wisdom" to anyone with a similar problem.

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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