My first western....advice please - (Sep/12/2008 )

Hello!

Im doing my first western next week, its for JunB. Today the other PhD showed me where the stuff was and I wrote some notes and hope I will remember what she said. Shes not going to be in next week so I guess ill try to do it by myself and im kinda nervous.

Ive been reading about aging gels and I wondered if someone could tell me how they would keep a gel moist overnight? Im going to be doing a big one in glass plates held together with clips. She told me to let it polymerize for three hours.

Uhm.any other useful tips?

I would recommend you to use freshly prepared gels (after about 1.5-2 hrs), I have been doing WB this way for 3 years and it works fine

You can keep it moist if your wrap it (after it has polymerized 3h at 37°C or RT) in several layer of wet paper tissues, so that every side of the glass plates are covered and no air can come in to dry the gel. Then put it in a plastic bag and seal it, so that the moisture won't get out. This way it is possible to store gels up to one week, but not longer, otherwise they won't give you good results anymore.

Using fresh ones is the best way though.

I am keeping my gels at 4C after wrapping with wet paper towels and sealing into plastic bags.

You can keep it moist if your wrap it (after it has polymerized 3h at 37°C or RT) in several layer of wet paper tissues, so that every side of the glass plates are covered and no air can come in to dry the gel. Then put it in a plastic bag and seal it, so that the moisture won't get out. This way it is possible to store gels up to one week, but not longer, otherwise they won't give you good results anymore.

Using fresh ones is the best way though.