improving terminator incorporation

Michael C. Gorry mcgorry at med.pitt.edu
Sat Nov 2 12:26:10 EST 1996


Hello,

I have been trying to improve the number of bases that can be read
on the 373 sequencer.  One of the problems that needs to be addressed
is to increase the incorporation of terminators past the 400 base mark.
I have been trying several methods (diluting the terminators, increasing
the number of cycles,increasing taq-FS,and adding DMSO.)  I have been unable
to increase signal strength to any significant amount with any of these methods.

Can anyone offer any other suggestions that would improve terminator
incorporation at longer points?  I have considered doing 4 set-ups, one
for each nucleotide.  This might use up too much DNA.  Has anyone tried
it in the past?

Any answers will be appreciated.

Thanks

Mike

-- 
Michael Gorry
mcgorry at med.pitt.edu




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