10 Hybridization Methods (Southern and Northern Blotting)
147
Subprotocol 7
Membrane Stripping
Procedure
Probes can be removed from hybridized membranes when rehybridization
is desired. Membranes must never be allowed to dry during or after hybridization and washing. Many protocols have been described for membrane
stripping. A simple one is the following:
1. Boil a solution of 0.1 % SDS.
2. Place the membrane in a glass tray. Pour the solution on the membrane.
Allow to cool to room temperature.
3. Check the membrane with a Geiger counter to monitor a decrease in
activity.
Note: Filtersthat have been stored for a long time (5 half-lives) of 32 P have
less than 5% of their initial activity. They may be used without stripping
after the absence of signal has been documented by exposure for an appropriate time.
References
Harnes BD, Riggins SJ (ed.) (1985) Nucleic acid hybridisation: a practical approach. IRL
Press, Oxford New York Tokyo
Southern, EM (1975) Detection of specific sequences among DNA fragments separated
by agarase gel electrophoresis. J Mol Biol 98:503-517
Membrane
stripping