Overview	RNA is transferred from gel to nylon membrane using vacuum gel transfer system.
Duration	2 hrs
2.1	Cut a nylon membrane about (or bigger than) the size of the denaturing RNA gel. Cut a filter paper with the same size as the nylon membrane.
2.2	Rinse the RNA gel with H2O.
2.3	Fill the wells of the RNA gel with melted agarose.
2.4	Wet the nylon membrane and filter paper first in H2O and then 10× SSC buffer
2.5	Put wet filter paper on the vacuum porous stage and make sure the filter paper is in the area where the cut window of the green plastic gasket is going to be. Place the wetted nylon membrane on top of the filter paper. Make sure there is no air bubble between membrane and filter.
2.6	Wet the seal o-ring on the base unit with H2O. Place the plastic gasket on top of the membrane/filter paper. Make sure the gasket covers the seal o-ring while the membrane/filter paper overlaps with the window of the gasket.
2.7	Gently place the gel on top of the gasket with the well-side up. Also make sure the gel overlaps with the gasket by at least 5 mm. Remove all the air bubbles between gel and the nylon membrane.
2.8	Place the sealing frame on top of the vacuum stage and lock it.
2.9	Start the vacuum source and adjust the pressure to 5 inches of Hg. Press the gel and along the window gently to apply extra pressure to help the vacuum sealing.
2.10	Gently pour 1 L of 10× SSC buffer into the reservoir. Place the lid on and transfer for 90 mins at 5 inches of Hg. Occasionally check the buffer level to make sure it is above the gel.
2.11	When the transfer is over, remove the sealing frame and drain the buffer. Remove the gel and take out the nylon membrane. Dry the nylon membrane between two sheets of filter paper.
2.12	UV crosslink the membrane twice to fix the RNA on the membrane and use a fine marker to mark the edge of the side with RNA.
2.13	Clean the gel transfer system thoroughly by rinsing with plenty of H2O.
Tip	After the gel transfer, the gel area inside the window of green gasket should be half as thick as the gel outside the window.
Tip	Gel can be illuminated with UV to check whether there is any remaining RNA.