Sequencing

Joseph C. Bagshaw jbagshaw at wpi.edu
Sat Nov 20 07:49:21 EST 1999


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.

********************  HAVE GENES, WILL TRAVEL  ********************
Joe Bagshaw, Worcester Polytechnic Institute
jbagshaw at wpi.edu
Roadkill on the information superhighway.

On Tue, 16 Nov 1999, Dr Raja Kota wrote:

> Hi Netters,
> 
>    Whats the best way to sequence a fragment which is over 1500 bp in
> length and is cloned into a vector.  Any references shall be appreciated.
> 
> Thanks in advance,
> 
> Raja
> 
> 





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