Hybridization buffer [for 4 probes] ddH20 25.0 mL 20X SSPE 12.0 mL 100X Denhardt's solution 2.0 mL 20% SDS 1.0 mL
Salmon sperm DNA [10 mg/mL; boil for 5 minutes and set on ice 5 minutes before adding]
0.2 mL dextran sulfate 2.0 g TOTAL 40.2 mL
- Replace prehybridization buffer in each hybridization bottle with 10 mL of hybridization buffer.
- Add probe to hybridization bottle. NOTE: do not allow the probe to come in direct contact with the membrane when first adding the probe. Make sure the probe goes directly into the hybridization solution in the bottle.
- Allow probes to hybridize for 18–24 hours at 65°C.