Sequencing from pLAFR3

Tsaltas, Dimitrios d.tsaltas at ic.ac.uk
Mon Apr 23 08:10:44 EST 2001


We are trying to sequence genomic clones on pLAFR3:

Anybody with any good protocols on extracting low copy plasmids and
sequencing their insert?

We are using the kits from Qiagen and then on sequencing we just increase
the number of cycles to 100 (of course we add more primer)
The amount of DNA we use is around 10micro garms. With less we don not have
any results.
With this protocol we take maximum 150-200bp sequence if it works.

Any sugestions?

Many thanks
Dimitris

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