We tried Accutrac and were not happy with the results. We found that it is
distinct lane marker while the gel is running but becomes a single blue
band across the width of the gel after the gel is analyzed, making it
useless for either automatic or manual lane tracking. We tried it with 5%
Long Ranger gels, with 36 and 48 cm plates, with TBE, with TTE, with 64
lanes and 96 lanes, with sharkstooth combs and porous combs and both with
and without the water loading protocol. All of these conditions had poor
results with Accutrac. We had better results using a labeled primer that
we designed for lane tracking, but still not optimal results. What were
your run conditions with Accutrac? Could you send some pictures of the
results? Thanks.
- George
At 09:36 AM 5/10/99 -0600, you wrote:
>We have been using AccuTrac for a couple of weeks now to help track our 96
>lane gels. It really helps define the lanes for manual tracking or
>adjustment of automated tracking, especially when more than a few lanes drop
>out. I don't see any indication that it helps with automated tracking yet.
>We're going to send some data back to CBI to help them refine the marker. We
>resuspend our samples in 3ul of marker dye and load 1ul. We load the marker
>dye in every well of a 96 lane gel, loading odd and even lanes 3 minutes
>apart. You could just use the marker in odd OR even lanes, but it works out
>to be relatively inexpensive per well so we define all 96 lanes. It's worth
>the cost just to be sure of lane tracking in a core laboratory where you
>have the potential for multiple lane drop outs. Give it a try!
>
>Margaret Robertson
>Director, DNA Sequencing Facility
>University of Utah,
>4A 432A, SOM
>50 North Medical Drive,
>Salt Lake City, UT 84132
>Tel: 581-4736
>Fax: 585-2978
>margaret.robertson@hci.utah.edu
>http://www.hci.utah.edu/groups/sequencing
>
>
>
>
___________________________________________________________
George Grills
Director
DNA Sequencing, Oligonucleotide, and GeneChip Microarray Facilities
Albert Einstein College of Medicine
713 Ullmann Building
1300 Morris Park Avenue
Bronx, New York 10461-1602
Tel: (718) 430-2657
Fax: (718) 430-8778
E-mail: grills@aecom.yu.edu
DNA Sequencing: http://leper1.ca.aecom.yu.edu/dnacore
Oligonucleotide: http://sequence.aecom.yu.edu/oligo
___________________________________________________________