RapidHyb protocol
Dominik Haudenschild - May 6, 1997 at Genzyme

Using random primed double stranded DNA probe (1/2 of 25ng reaction = 2ng/ml) or RNA probes (1/2 of standard reaction = 6ng/ml)

• Prehyb HyBond N+ for 15 minutes in RapidHyb at 65°C (70°C for RNA probes)
If using roller bottle in hyb oven, use at least 10ml
• Boil probe for 2-5 minutes at 95°C to 100°C
• Snap-cool probe on ice
• Add 1ml prehyb solution to probe, then add probe to membranes
• Hybridize at 65°C (70° for RNA probes) for 1 to 2 hours
Overnight is OK but background increases
• Wash once 20 minutes at room temerature with 5x SSC + 0.1% SDS
• Wash twice 15 minutes at 55°C to 65°C with (1 to 0.1)x SSC + 0.1% SDS
• Wrap in Saran wrap and expose to film overnight