QuickHyb protocol

Dominik Haudenschild - May 6,1997, at Genzyme

Using double stranded DNA probe, random primed
• Prehyb membrane in 35µl/cm2 of QuickHyb at 68°C
If using roller bottle in hyb oven, use at least 12ml
• Add 100µl of sonicated salmon sperm DNA to probe
• Boil probe for 5 minutes at 95°C
• Add 1ml prehyb solution to probe, then add probe to membranes
• Hybridize at 65°C for 1 to 2 hours, overnight is OK but background increases
• Wash twice 15 minutes at room temerature with 2x SSC + 0.1% SDS
• Wash once 15 minutes at 55°C with 0.2x SSC + 0.1% SDS
• Wrap in Saran wrap and expose to film overnight