VERTICAL DGGE GEL - (Apr/12/2006 )
can anyone plese tell me how to prepare a vertical DGGE gel ??????? and also how to pur it
can it be done without formamide?
Protocol DGGE ( got it by goggle search )
i got that proocol and others from my serach on yahoo but i cant do it ....seems that the devics im
working on doesnt have what the all call the gradient chamber ?
i was woundering can i pour first he high conc gel then immediatly the lower on to fill to the ends
of the glass chamber ????
that i a half gel ( lower part with high conc and upper part ( upper half with lower conc )???????
another thing also is formamide a must ? whats its replacemnet ?
We do some in our lab, and from what our DGGE specialist tells me, the answer to both your questions are No. You must find a gradient mixer (Fisher sells some), because otherwise you are not going to have anything that look like a good pattern. And she doesn't know anything that can replace formamide. Sorry.
thanks alot for replying ,,,,,i wll do m y best to look into the situation
I used to use a BIORAD system. They have a pretty low tech gradient mixer that is easy to use and to clean. I couldn't find the catalog entry for it on the biorad site but i found a description of their DGGE sytem here
Their gradient tool is simply a wheel that you attach two 50 ml syringes-with tiny hoses on them, that are filled with high and low concentration polyacrylamide solutions.
The trick is to dilute your TEMED a bit more than recommended in the protocol, so that it doesn't polymerize while you are still trying to pour it. The gels set a bit slower but it helps , until you can work faster.
P.S. It was right infront of me and i didn't see it before- here