DNA Hybridization - Removal of Bound DNA


Jeff Lawrence

1. Folowing autoradiography, DO NOT allow membranesto dry. Otherwise, DNA is irreversibly bound.

2. Place membrane in 100 mL of: 
	400 mM	NaOH	8 mL	5 N NaOH
	0.1 %	SDS	1 mL	10% SDS
			91 mL	ddH2O
3. Incubate at 45° for 30 min.

4. Decant solution. PLace membrane in 100 mL of: 
	0.1 X	SSC	5 mL	20X SSC
	0.1 %	SDS	10 mL	10 % SDS
	0.2 M	Tris, pH 7.5	200 mL	1 M Tris, pH 7.5
				785 mL	ddH2O
5. Decant solution. Rinse filter in water. Store.