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southern blotting also known as southern hybridization is a technique used for the detection and quantification of specific dna levels using a hybridization dna probe southern blotting makes use of the gel electrophoresis method the steps involved in southern blotting are as follows isolation or separation of dna the isolation process is done by extraction afterward the purify genomic or plasmid dna is partially digested by a restriction enzyme restriction endonucleases these enzymes cut double-stranded nucleic acids at certain sequences the resulting mixtures of different sized linear fragments are then loaded side by side into wells formed in an agarose gel separation is done ing to size after this the dna is now negatively charged the dna is then transferred to a solid membrane the membrane driven by capillary forces is exposed to intense ultraviolet uv light to induce dna cross linking since the dna fragments are double-stranded they have to first be denatured by pre-treatm