You can help by adding to it.
Cut off the top left corner of the membrane. This edge will serve to orient the membrane. The paper towels should be slightly larger than the gel. Place a plastic wrap around the Whatman papers to prevent short-circuiting.
Remove the air bubbles by gently rolling over the stack with a disposable 5-ml pipette. Place the membrane on the top filter paper.
Remove the bubbles by rolling a pipette over the surface of the membrane. Lay the gel onto the membrane by starting at one side of the stack.
Take care to observe the buffer flow below the gel as the gel is placed onto the stack. In case air bubbles are trapped between the gel and the membrane, lift the gel to remove the air bubbles.
Remove the air bubbles.
The use of two reservoirs is important to prevent distortion of bands during the transfer. Place a glass plate on top of the assembly. Cover the assembly with a cling film to minimize evaporation. For convenience, we leave the transfer overnight.
Disassemble the transfer unit using forceps.
Do not let the membrane dry before cross-linking. The conditions for the UV exposure are critical. Too short exposures will result in inefficient cross-linking of the RNA, and too long exposures or exposure of dried membranes will result in too strong cross-linking, decreasing significantly the sensitivity of the hybridization steps.Filter papers come in a broad range of glass fiber, qualitative and quantitative types for all needs.
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