Thanks to everyone who has helped me so far with this problem. I thought I
would post a quick summary of some of the suggestions I have been given and
some of the results I have had myself.
Basically the problem was that after having our 373 and 377 ABI machines
upgraded to take 48 and 60 well square combs we wanted to find a way to
protect the wells from collapse. Our first attempts using bind silane gave
beautiful wells but also almost welded the acrylamide to the glass plates.
We have a high through put of gels daily and this caused problems with
washing.
Bind silane binds to glass plates and, because it contains acrylamide
monomers, when the poured gel polymerises it binds to the silane via these
acrylamide monomers so sticking the wells to the plate.
A couple of recipes were posted to me
1.5ul Bind silane
700ul Ethanol
160ul 10% acetic acid
Wipe onto the plate with a tissue - only need to repeat after 2-3 gels
17ul Bind silane
170ul 10% acetic acid
10ml ethanol
wipe on with a cotton bud.
200ml dH20 adjusted to pH 3.5 with Acetic acid plus 800ul bind silane, stir
till clear then dip the top of the plain plate into the solution for 10-60
minutes, rinse with dH20
I have tried these three recipes and they all give me great wells but still
stick the acrylamide to the glass.
The removal of the acrylamide - suggestions were to scrape off the gel with
a razor blade (gulp), or to soak in 0.1M HCl or NaOH. For us this is not
feasable due to the large number of gels poured daily.
I'm going to carry on looking into this and will post my results if and
when I succeed.
Thanks again to everyone who replied to me and I hope this helps someone
else in the future.
Alison Brown
