LongRanger on Automated Sequencer?

Michael Gorry mcgorry at med.pitt.edu
Wed Aug 9 19:12:52 EST 1995


In article <1995Aug3.175425.328 at immunex.wa.com>,
hollingswor at immunex.wa.com wrote:

> The recent question about gradient gels on an ABI373 prompted this question.
> 
> Has anyone tried Long Ranger gels?  What percentages, running parameters, 
> matrix/mobility concerns?  How were the results and lenth of read?
> Thanks
> Tamy Hollingsworth

I have tried using longranger gels on the ABI 373.  While a 5% gel gave me
fantastic results,
by this I mean longer reads and sharper peaks, there were several
problems.  The biggest
one was the fact that the 373 is incabable of calling bases that move
across the scanner at
rates different from the default.  This meant that I had to manually go
through the entire
sequence results and correct any bases that weren't called (about every
third base). By lowering
the power to 20 watts, I got a better calling but the gel took
considerably longer.  The gel is better
than a regular polyacrylamide gel and will give good base calls out to 600
bases without trouble.
I recently contacted ABI to see if they are going correct for this, and
while the new 377 can, it
doesn't look as if the old 373 has any chance.  There was no problem with
the matrix.

-- 
Michael Gorry
mcgorry at med.pitt.edu



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