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southern blood is a classic technique in molecular biology that reveals information about dna identity size and abundance and it can be used for detection of a specific dna sequence in dna samples the first step in southern blotting is the digestion of the dna samples with an appropriate restriction enzyme a restriction enzyme also called restriction endonuclease is an enzyme that cleaves dna into fragments at or near specific recognition sites within molecules known as restriction sites once the restriction enzyme is added the samples are incubated at 37 degrees celsius overnight restriction enzymes recognize a specific sequence of nucleotides and produce a double-stranded cut in the dna therefore various dna fragments of different sizes are obtained then the dna fragments produced by restriction endonuclease digestion are separated by gel electrophoresis for gel electrophoresis separation a loading buffer is added to the dna samples and it is used as a tracking die which migrates in