Post Hybridization

1. Remove the membranes from boxes.
2. Wash the membranes with 1 L 0.25X SSC 0.2 % SDS (per 15 membranes) on a shaker three times at room temperature for 5 min each time.
3. Wash the membranes with 1.0 L 0.25X SSC 0.2 % SDS (per 15 membranes) twice at 60 C in a shaking water bath for 30 min each time.
4. Remove the membranes and place on 3 MM paper. Remove excess solution with paper towel briefly. From now on, do not let the membranes dry completely as the counts will be bound irreversibly.
5. Place the moist membranes on a plexiglass sheet and cover membranes with Saran wrap. (Do not use Reynolds wrap! It makes membranes wrinkle)
6. Place x-ray cassette. Cover with a sheet of x-ray film and one intensifying screen. Store at -70 C overnight.
7. Develop the x-ray film and label. Store the wrapped membranes at 4 C. Counts should be removed at least within 2 days.