Skip- We had a bad run of distorted mobilities and disappearring bands several
years ago. So here are my comments from that experience-
I tried changing plates to get rid of the problem several times, but it would
eventually reappear. Once, I had the problem show up on a set of plates on
its second use. So new plates are no guarantee.
Like you, we are a small lab without a dishwasher. I wash plates in Alconox
(scrubbing with a gloved hand, not a sponge) and rinse the plates copiously in
hot tap water (prop the plate in the sink and let the water run for 5
minutes), then copiously in room-temperature dH2O. Never touch clean or dirty
plates with bare hands or Kimwipes. Don't stand them to drain with the edge
against labmat or anything but a clean plastic rack. Allow to air-dry.
During the time when I was having a lot of gel problems, I was using Alconox
but rinsing less fanatically, draining the plates on labmat, and occasionally
touching them with clean ungloved hands. Since changing my plate-handling
routine I haven't seen the disappearring bands for over three years and have
only changed plates due to breakage. I never acid-wash them.
Personally, I believe properly cleaned plates could be used indefinitely. The
trick is to develop a consistent routine that works in your lab with your
personnel, your available space, and your water supply.
I have only used ABI plates but in view of the cost I'd like to hear from
others who have tried alternative suppliers, too.
I know this problem is aggravating . Good luck! Regards,
Allison Pinder
DNA Sequencing Core Facility
Department of Embryology
Carnegie Institution of Washington
115 West University Parkway
Baltimore, MD 21210
410 554-1207
pinder@mail1.ciwemb.edu
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Dear fellow sequencers,
We're wondering if it's time replace our glass plates (377 standard,
36cm.). They've been used constantly for over 3 years. We are plagued
occasionally by mobility problems in some lanes and the "disappearing -
reappearing" bands. We have temporarily solved the problem by "acid/base"
washing the plates. We have used Multiterge, Citrinox, Alconox, and
Seque-soap to wash the plates, but the problem inevitably returns. (We
understand a very hot DI water rinse works well, but we can't afford a lab
dishwasher).
My question is, does age "predispose" plates to this problem or do proper
cleaning methods allow plates to be used indefinately?
We are considering buying plates from Sigma. They're half the price of
ABI's and their ad claims use by genome centers and core facilities
worldwide! Are they comparable to ABI's?
The latest issue of American Biotechnology Laboratory has an ad for a
product called GELPLATE-clean. It claims that this product removes the
contaminanting molecules on the glass surface without the need for soaking.
Does anyone have experience with this product? Does it leave any
fluorescent residue?
Any advice is welcomed.
Best regards,
Skip Vaught
DNA Sequencing Service
University of Arizona
THE DNA SEQUENCING SERVICE
UNIVERSITY OF ARIZONA
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Skip Vaught- jev@u.arizona.edu
Brian Coullahan- coullaha@u.arizona.edu
Mark Schwartz- schwartm@u.arizona.edu
Amy Raymond- raymonda@u.arizona.edu