How Much PCR product for sequencing?

Jenny Cassady cassady at biosci.uq.edu.au
Thu Apr 18 04:02:35 EST 1996

Dear Autosequencers,

We are in a bit of a dilemma - Does anyone have a good "rule of thumb"
for the amounts of PCR product required for sequencing, per 100bp of
product length? We are using Perkin Elmer Taq FS labelled terminator
chemistry. We have found a lot of the manuals were written for
ordinary Amplitaq. The PE kit instruction manual suggests different
amounts from their Comparative PCR sequencing booklet for the same
enzyme. Our users that are getting good direct sequencing results are
not confident of the accuracy of their estimations.  Confusing, hey?

Jennifer Cassady
DNA Sequence and Analysis Facility
The University of Queensland
St. Lucia, Qld. 4072

Ph:  +61-7 3365 4601
Fax: +61-7 3365 4388
email : cassady at biosci.uq.oz.au

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