khatipovNO at NOuchicago.edu
Fri Oct 26 17:46:57 EST 2001
It has been discussed before, as well as it is mentioned somewhere in
Bio-Rad's or other manuals that cracks are due to acetic acid in the gel and
air bubbles. I generally run same precast gels as yours, stain/fix them in
0.1% Coomassie R250 : 10% acetic acid : 40% ethanol for at least 1 h
(overnight OK), destain in 10% acetic acid : 40% ethanol followed by water
(not necessarily distilled). Quite often I would destain with distilled
water in a microwave (boil 2-3 min). I dry the gels in a gel drying
cassette/frame (not in a dryer) between 2 sheets of cellophane (e.g.,
Promega's Gel Drying Film). I hope you know how to use gel drying frames, so
I will not go into details here. Important thing is to remove ALL air
bubbles between the sheets of cellophane - literally put your gel in a pond
of water and cover with the second cellophane sheet, and then squeeze or
drip extra water out. It is better not to use small frames. 20 x 30 cm ones
per 1 10 cm gel worked best in my hands.
Let me know if that worked.
"Terry Young" <tlyoung at u.washington.edu> wrote in message
news:20c8e91.0110261357.144cb5f8 at posting.google.com...
> We are using Biorad's Protean II Ready Gels to seperate protein
> translation products.
> These are the 8-16% (and 10-20%) gels...the problem is they often
> crack into beautiful mosiacs when we removed them from the gel
> dryer...that give lousey results.
> We fix our gels in 50% methanol, 10% glacial acetic acid, 38% water,
> 2% glycerol.
> We have tried several different drying conditions, including 60 C for
> 2 hrs, down to 50 C for 1 hour and even overnight drying witnout heat.
> We have also tried various concentrations of glycerol to no avail.
> It seems that we cannot consistently get dried gels without
> cracks...and these are often too bad to be able to interprete the
> Has anyone already been through this???
> Terry and Stephanie
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