abrf@aecom.yu.edu
Subject: Re: DNA sequencing
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Hi, Susan,
We have had a Beckman-Coulter CEQ2000XL for a few weeks now and have
been making good use of it. We would be happy to communicate about
trouble shooting, etc. with you and others.
Our experiences have been generally positive; the sequencer seems easy
to operate.
Some trouble shooting items that we have learned in the short time we
have been using it are:
The chemistry protocols in the manual work well, especially if you
follow them exactly as stated.
100 nanograms of template for a typical plasmid construct (approximately
3 to 5 kb total size) seems about right most of the time.
If the start of the data is delayed from the norm of about 20 minutes
into a run (for LFR-s) and the data doesn't go past 600 bases, try less
template in the reaction. The current profile is also a useful indicator
of too much template.
Some of our users bring us template with high G/C content and we have
been trying ideas not mentioned by Beckman-Coulter. Some of these have
been successful, but we need more experience before we can say what
works best.
Ron Redder
-- Genomics Core Research Facility Center for Biotechnology N319 The Beadle Center University of Nebraska Lincoln, NE 68588-0665 Tel: 402-472-5453 Fax: 402-472-3139 dnaseq@unlnotes.unl.edu
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