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.
Last Update: Thursday June 19 2014
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Eric Kofoid
eckofoid at ucdavis.edu