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in this next step we will transfer our separated proteins out of the gel and into a solid membrane or blot this is based upon the same principle as the previous step in which an electric field is applied to move the negatively charged proteins towards a positive electrode transfer can occur under wet or semi-dry conditions here we will demonstrate the traditional wet transfer method start by removing the gel from its cassette cutting off the top portion containing the Welles notch the top left corner to indicate gel orientation float the gel and transfer buffer while preparing the transfer sandwich to make the transfer sandwich you will need a cassette sponges filter paper the gel and your choice of either PVDF or nitrocellulose membrane PVDF must first be activated by soaking the membrane and methanol for 2 minutes but other than this the choice of nitrocellulose or PVDF membranes is a personal preference notch the top left corner to indicate blot orientation and incubate memb
