David L. Haviland, Ph.D.
HAVILAND at KIDS.WUSTL.EDU
Fri Sep 9 13:07:09 EST 1994
In <34fiie$9h2 at mserv1.dl.ac.uk> JAB5 at VAX.YORK.AC.UK writes:
> In addition to the current PEG thread, it does seem to be important
> to remove all PEG from ssDNA template prepared for sequencing. I can
> only vouch for this from using Klenow for seq so I'm not sure about
> T7/Sequenase or cycle methods.
I'm not sure it's a debate about PEG, PEG works but certain rules must be
I can vouge for T7. In the past we used M13 for cDNA sequencing and used
USB Sequenase T7 for single stranded sequencing. If the PEG was not
removed - there was NO sequence. No "maybes", no partial sequence - nada!
There is something about PEG that completely inhibits the function of
enzymes, Taq Pol and T7 are what I have experience with...
Hope this helps,
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