Yesterday I ran an PAGE gel to detect miRNA. All was going well, then I set-up the apparatus for the transfer. I did it as any capillary protocol says: dish witn 20x SSC, glass over support, wet paper over glass, gel over paper surrounded with parafilm, membrane over gel, wet paper over membrane, paper towels and a weight. However, prior to using the membrane I did not wet it (this is where the oops comes in). How important is it to wet the membrane? Would I get any transfer if I failed to do this?
As I type this I am preparing a second membrane, hopefully it will be OK.
I used a Millipore Immobilon Ny+, in case you were wondering.
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Northern blot: Wetting a membrane
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