Double stranded DNA sequencing

Susan L Forsburg susan_forsburg at
Wed Mar 16 16:17:22 EST 1994

>: I am currently attempting to sequence double stranded plasmid DNA
>: minipreps using the USB sequenase 2.0 Kit.  Unfortunatly after
trying several
>: DNA isolation methods that I could think of or find I am not
having good
>: results in that > 70% of my reactions produce either no bands or
bands in all
>: four lanes all the way up the gel.  I am interested in a miniprep
DNA isolation
>: technique (Kit or otherwise) that reliably yields 50% or greater
>: sequencing reactions.
>: Bob Burns

I have always had difficulaty with "standard" ds sequencing. 
Recently I had excellent luck using a PCR based sequencing kit from
USB called Delta-Taq cycle sequencing, with cycled labelling
protocol.  The sequence looked like single stranded, completely and
gorgeously readable.

Usual disclaimers,


Susan L Forsburg
Molecular Biology and Virology Laboratory
The Salk Institute for Biological Studies
email:  susan_forsburg at
        formerly forsburg at

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