help sequence from puc

Randall J. Cohrs COHRS-R at Oberon.HSC.Colorado.edu
Wed Nov 9 11:11:05 EST 1994


> In article <carmean-0811941114350001 at creslab2.biol.sfu.ca>, carmean at sfu.ca
> (Dave Carmean) 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.
> > 
> > William D. Brown
> > Department of Biological Science
> > Simon Fraser University
> > Burnaby, B.C.  V5A 1S6
> > Canada
> 
> I use the Sequenase (USB Biochemicals) kit for sequencing double-stranded
> DNA from plasmid preps (mini or regular) with great success. Could be your
> preps aren't quite clean enough, in which case RNase treatment might
> help.  
> bb (beckthol at acs.ucalgary.ca)
> 
 - - -

We also use sequenase with same Great success, only thing we check is 
primer to template ratio of 1.6 pmole to 2.0 pmoles



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