VERTICAL DGGE GEL - (Apr/12/2006 )

hi

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 )

gud luck

hi

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 ?

thanks

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.

hi

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.
good luck
P.S. It was right infront of me and i didn't see it before- here
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