Southern blot - (Sep/25/2008 )
My problem consists on the use a pure nitrocellulose membrane in a southern blot test. After an over nignt tranfert of DNA from an agarose gel via SSC 20x, the membrane becomes very weak. When i try to put it in the bottle of hybridazition, it brea down.
please is there some one who can give me the composition of the transfert buffer of a nitrocellulose membrane?
that is the problem with nitrocellulose. it gets brittle after wetting and drying. otherwise, ssc is compatible with nitrocellulose.
we use supported nitrocellulose or nylon (neutral or charged) for southern blots.
if you have to use unsupported nitrocellulose then you will have to handle carefully (maybe rewet before putting into bottle).