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in this video we will show you how to set up a polyacrylamide gel for wet tank transfer in a mini transplot cell after you have removed your gel from the gel cassette equilibrate the gel in a tray with transfer buffer for 15 minutes on a rocking platform pre-soak blotting paper and fiber pads in the transfer buffer and ensure complete coverage if using a nitrocellulose membrane pre-wet the membrane and transfer buffer if using a pbdf membrane pre-soak in 100 methanol to make the blotting sandwich place the gel holder cassette with the black side down and a tray with sufficient transfer buffer place one pre-wet fiber pad onto the black plastic followed by a pre-wet sheet of blotting paper take the gel and carefully place it onto the blotting paper being careful to avoid trapping excessive air bubbles now place your pre-soaked membrane onto the gel using a roller remove any air bubbles between the gel and the membrane to ensure maximum and even protein transfer place a second she
