What Is Involved in Southern Blot Hybridization

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The Answer is DNA samples, almost always genomic DNA samples, are treated with a suitable restriction endonuclease that cleaves the DNA into double-stranded fragments of a manageable size. The DNA fragments are separated according to size by agarose gel electrophoresis (DNA is negatively charged and will migrate towards the positive electrode through the porous agarose gel; small fragments move faster through the gel because there is less frictional resistance than with the larger fragments ).
After electrophoresis, the gel is immersed in strong alkaline solution and the resulting denatured DNA is transferred by capillary action to a nylon membrane that is placed in contact with the gel ("blotting"). The individual DNA fragments become immobilized on the membrane at positions which are a faithful record of the size separation achieved by agarose gel electrophoresis.
Subsequently, the immobilized single-stranded test sample DNA sequences are allowed to associate with labeled single-stranded probe DNA. The probe will bind only to highly related DNA sequences in the test sample DNA, and their position on the membrane can be related back to the original gel in order to estimate their size.

What Is Involved in Southern Blot Hybridization

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What Is Involved in Southern Blot Hybridization

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