373A sequencing of GC-rich templates

Kevin Clark kclark at immsvr.jr2.ox.ac.uk.
Wed Jul 6 11:59:30 EST 1994


We are having problems sequencing through a region of approx 200 bases
of almost total G's and C's. Does anyone have any experience of this
kind of template and how to get through it.

We can use dye primers or terminators and have access to Hybaid
Omnigene and Perkin Elmer 480 PCR machines.

The two most obvious options to try are adding DMSO and/or Triton X-100
to the PCR but does anybody have any experience of concentrations to
use etc.
The DNA is in M13.



\/\    /\/\    /\/\    /\/   kclark at immsvr.jr2.ox.ac.uk
 \/\  /\/\/\  /\/\/\  /\/    Institute of Molecular Medicine,
  \/\/\/  \/\/\/  \/\/\/     Oxford,UK.
   \/\/    \/\/    \/\/ I've been here for 5 years and they only    
                        hung me the right way up yesterday.



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