The hybridization of the Northern Blot Membrane
20-25 mL hybridization buffer (0.5 M Sodium phosphate buffer pH 7.2 containing 7% SDS) and the rotating bottle were heated in a hybridization oven (Hybrigene, #Z649570) at 62°C in darkness. The membrane was placed in the bottle and rotated for two hours. The fluorescent-labelled DNA oligonucleotide was added to 10 μM final concentration, and hybridization occurred overnight. The next day, the wash buffer (20 mM sodium phosphate buffer pH 7.2 containing 1% SDS) was warmed in a water bath to 43°C. The membrane was washed with this pre-warmed buffer in a temperature-controlled orbital shaker in a metal box, preventing light exposure. The membrane was washed thrice for 5 minutes with approximately 250 mL of the wash buffer at 43°C. Finally, the membrane was placed into a plastic envelope. The scanning of the membrane was done in the Amersham Typhoon™ laser scanner.