DNA Footprinting Gel - (Feb/13/2008 )

Hey,

Does anyone have any idea/tips on what the best way to get a sequencing gel (10% TBE/UREA acrylamide) off the glass plates. We are trying to do footprinting, and when it comes to transferring the gel off the plate, and onto paper to put it into the cassete it rips! Anyone have any ideas?

Thanks!

Elena

You can usually do this submerged in water or buffer -- the liquid supports the gel while it is loose.

And then try and put paper into the buffer underneath the gel?

Thanks for the idea, Ill definitely try it!

the way we did it:

both plates had been siliconized or rain-xed prior to pouring the gel.

after the run, split the plates, keeping the gel on one.

cover the gel with fixative (methanol/acetic acid/water).

overlay filter paper sheet (add more fixative as necessary), work bubbles out with glass rod.

blot off fixative with stacks of paper towels, completely covering filter paper, do not press.

when paper towels come off dry, then carefully lift the filter paper, peeling from one end to the other.

the gel should come off smoothly.

we then dried on vacuum gel dryer (covered with plastic wrap), placed in cassette and exposed.