Reply to: RE: Sequencing plates and plate cleaner
Skip,
I think eventually the plates are unusable. They seem to collect tiny =
knicks in the polished surface from all the washing/acid etc. I tried to =
prove to Christine a long time ago that these exposed knicks were the =
place that small nodules build up over a few gels and cause the "=
disappearing bands". I wanted to run more gels and look at the bumps under =
a microscope and she wanted to buy new plates. We bought new plates.
This etching might be the reason why the 3700 might never do as well as a =
slab gel. With a slab gel you get to use two highly polished surfaces to =
contain the migrating DNA sample. It is hard to polish the inside of a =
capillary. Same difficulty showed up in all the attempts to use etched =
microchannels in glass plates to contain the sample.
A quote from an abstract by Steve Swierkowski, Microchannel Process =
Development and Fabrication for DNA Sequencing =
..... explanations of technique for cleaning......then.....
"This special procedure was essential to reduce the microchannel etching =
defects by many orders of magnitude that would have otherwise rendered the =
plates useless for high resolution genome sequencing". =
see:
http://www.ornl.gov/hgmis/publicat/97santa/seqtech.html
Sounds like your plates. We buy currently from Sooner Sci.. Sigmas I'm =
sure will be fine also
hope this helps, Paul
Paul Morrison
Molecular Biology Core Facilities
Dana-Farber Cancer Institute
Jimmy Fund Two
44 Binney Street
Boston, MA 02115
paul_morrison@dfci.harvard.edu
http://mbcf.dfci.harvard.edu
617-632-3082
fax 632-4814
----------------------------
Skip Vaught wrote:
>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
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Reply to: RE: Sequencing plates and plate cleaner
Skip,