HELP with sequencing acc NAR 16(22):10915

David Johnson djohnson at acpub.duke.edu
Mon Aug 29 13:37:16 EST 1994


Greetings!

Has anyone used the protocol in Nucleic Acids Research
16(22):10915 (1988) ???

In summary, a single-stranded (M13) template was
sequenced using Taq polymerase, in which 7-deaza dGTP
and radiolabeled nucleotide were used in a reaction 
at 70C. That's right, the M13 17-mer universal primer
(-20), which has a Tm of approx 52 C, is used in an 
extension reaction with M13 template and Taq pol
WITHOUT AN ANNEALING STEP !!

How can the primer anneal?

Has anyone used this method successfully?

I am particularly interested in an adaptation of the
method for double-stranded templates (namely, PCR
products).

Please send responses via email !! (since I don't
read the news here very regularly).  THANKS !!!

Dave.

-- 
David W. Johnson                  Power is no substitute for skill
Amateur Extra WA4NID              QRP ARCI 6546
email: djohnson at acpub.duke.edu    G-QRP 4864
packet WA4NID at KB4WGA.NC.USA.NA    NorCal 355



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