Hybridization of Digoxigenin-Labeled Probes to Southern Blots and Detection by Chemiluminescence

Probes prepared with either digoxigenin- or biotin-modified nucleotides can be hybridized to Southern blots to detect target nucleic acid sequences. These methods offer an attractive alternative to “radioactively tagged“ probes in terms of safety, cost, and efficiency. Most previous nonradioactive strategies utilized the detection of the modified base by the use of a coupled antibody- or avidin-alkaline phosphatase. The blot with the bound alkaline phosphatase was then treated with a compound, such as Nitro Blue Tetrazolium (NBT), that was converted to an insoluble, colored compound at the site of hybridization, thus facilitating visualization of the hybridized probe.