problem in separating gel - (Jul/20/2007 )
when I prepare the separating gel I put all component and put it in the two glass and cover the surface by isopropyl alcohol
after while the polymerization occur but in the lower part only even i left it for 1 hour so when I pour 5 ml gel it give me 3 ml gel only what is the wrong in my component
does anyone work in identification of meat according to species by PAGE elecrophoresis?
please communicate with me in this research
thanks
Did you ever try to cover it with water?
no because the protocol say using isopropyl alcohol to remove pubbles and isolate the gel surface from air to help polymerization better
and i try this method several time and it is succed but my problem appear recently
Such a differencecome surely from sthg wrong in your apparatus. I admit that on minigel the surface slow from 8cm to 7,5 at most.
Check your apparatus by filling with water only, wait 15' and see the level.
Isopropanol dessicates the srface of the gel too. So don't wait 1h.
after while the polymerization occur but in the lower part only even i left it for 1 hour so when I pour 5 ml gel it give me 3 ml gel only what is the wrong in my component
Hi There,
do you put all the reagents into a tube and mix well before you pour the gel? Also try to make APS fresh. the older APS gets, the longer gels take to polymerise. Store your prepared APS at 4C. Good luck!
thanks for all for reply
actually APS is prepared from 4 months and stored in freezer
and my supervisor is in hospital and there is no solution to get fresh APS
so what can I do with my APS
Instead of isopropanol try use water-saturated butanol. I don't have problem with this one even when I leave gel for overnight polymerization.
Degassing of your gel solution may help a bit.
Are there not other people in your lab to give you APS? Or maybe all you need is to look around shelves?