Sequencing

Tom Knight tk at pasteur.ai.mit.edu
Sat Nov 20 11:23:49 EST 1999


"Joseph C. Bagshaw" <jbagshaw at wpi.edu> writes:

> 
> I've tried several approaches and the one I favor is primer walking.
> Sequence as far as you can with the flanking primers in the vector, then
> design custom primers and go further.  Given the price of custom oligos, a
> primer will cost you no more than two or three hours of your lab tech's
> time costs.  With an insert of about 1500 bp you should be able to
> sequence both strands with just a few custom primers.  We sequenced an
> insert of nearly 9 kb by primer walking.

Also, you end up with a set of probably useful primers to the sequence,
which, presumably, you are interested in else you would not be sequencing it.
If you are clever about the choice of your primers during sequencing,
these can be placed in strategic locations for future PCR creation of
hybridization probes, e.g.





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