HELP with sequencing acc NAR 16(22):10915

David Johnson djohnson at
Mon Aug 29 13:37:16 EST 1994


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

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

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


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

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