Stripping DNA probe from dna dot blot - (May/23/2006 )
i need to to strip my probe (DIG labeled) from a dot blot. i have used the method in the DIG handbook (0.2 NaOH, 0.1% SDS), 37 degrees 30 minutes. it didn't seem to work well and i suspect reason is this only removes the alkaline-labile DIG label rather than the probe itself. is this correct?
i need to remove the DNA itself, as my subsequent probe recognuzes the same sites ( i am doing differential screening). any one have a good method (seen several on the net and dont know which to choose...)
i am using a Nytran supercharge Nylon membrane (Schelicher and Schull)
i boil 0.5%SDS and add it on the membrane gentle swirl and let the stuff cool to RT.
And that's ok to remove DNA probes.