help sequence from puc

Klaus Salger salger at
Wed Nov 9 17:45:50 EST 1994

Dave Carmean (carmean at wrote:
: I would like to receive copies of protocals for standard sequencing of
: double-stranded DNA.  I am having difficultlies getting good sequence from
: puc and I'd like to compare your methods with ours to help pinpoint the
: problem.

I use the fmol sequencing kit from Promega with direct incorporation
of 33P-ATP. This gave me sequences (350bp) with plasmid preps that
were almost impossible to sequence with Sequenase. I find cycle sequencing
to be very convenient, put reagents together and place in the cycler,
that's all.
Hope this helps
Klaus Salger                phone : ++49 (0)89 5902 -502
Zoologisches Institut       FAX   :                 -450
AG MacWilliams              e-mail: salger at
Luisenstr. 14
80333 Muenchen

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