2 replies to this topic

[shabna]

pito, on 25 April 2009 - 07:48 AM, said:

Anyone who reads this topic or needs info on the subject, be sure check the files attached.

Hi everybody,

In SDS-PAGE, my separating solution is not fully gelled. Everytime i get 200 to 500ul of extra solution without gelled. i would like to know what is reason for this and how to correct it.

[bob1]

How do you mean?  You make up a working solution, add APS and TEMED, then pour the gel and a little bit doesn't polymerise?  If so this may be because your gels are not de-gassed and O2 inhibits the polymerisation of acrylamide.  Or it could mean that your APS is a bit old, try making some fresh solutions.  APS will keep in the freezer for about 6 months.

[kingswill]

What do you mean by extra? Is it the solution left behind after you pour your gel? Then is your gel ok? If you gel is ok it's not a problem I suppose?

If you gel is not set, first make sure you mix the solution gently to minimize air in your solution (as bob stated the reason), or you may go for de-gasing (Never done this before but if gas is a real problem) . Then check your APS (this is the most likely part that causing un-set gel), prepare a new 10%APS if needed. Once I found my APS powder looked hydrated and my gel didn't set even I prepared APS solution fresh. I borrowed APS powder from other and it worked fine. Then check Temed and Acrylamide.