The wells of my gel break from one corner even before the gel polymerizes. Can anyone provide some tips and/or possible solutions to this issue? I have to separate DNA using PAGE.
How does your gel recipe look like? How much APS and TEMED do you use?
Greeting Mr. Praetorius
For the 6% acrylamide gel, I have mixed the following ingredients
1. 10X TBE = 2.5 ml
2. 30% acrylamide stock = 10 ml
3. APS = 350 µl
4.TEMED = 50 µl
5.dH2O = 37.1 ml.
Break, or never form? This is odd!
- the wells are the last to polymerize- give it a little longer.
- Try dropping APS and TEMED to make it polymerize slower.
- Is there enough solution left during polymerization? Add a bit of solution along the comb so if the gel shrinks, more solution can be sucked in.
- maybe base bath your plates in case there is some odd residue causing problems for you.
Good luck!
Polymerization of Acrylamid is highly oxygen sensitive. Try to minimize the contact area of solution and the atmosphere. You can also try to degas the solution before you add APS and TEMED. In some gel casting systems the surface area exposed to the atmosphere is quite big and can cause trouble.
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