SOUTHERN BLOT Capillary Transfer of DNA to a Membrane. ABE WORKSHOP JUNE 6-24, 2005. Purpose. Detection of specific sequences among DNA fragments separated by gel electrophoresis. Restriction fragments appear as a smear rather than as discrete bands on the gel.
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JUNE 6-24, 2005
Preparing probe: Combine 10 µl template DNA & 6 µl dH2O. Denature the DNA by heating in a boiling water bath for 10 minutes and quickly chilling on ice. Mix DIG-High Prime thoroughly and add 4 µl to the denatured DNA, mix & centrifuge briefly. Incubate for 1 hour at 37ºC. Stop the reaction by adding 2 µl 0.5M EDTA.