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Nicole Hernandez

Southern Blotting is a technique utilized in molecular biology for the purpose of detecting a certain DNA sequence in DNA samples. British biologist, Edwin Southern, invented the method in the mid-1970s. The method of Southern Blotting involves the transferring of DNA fragments (separated by gel electrophoresis) and being placed upon a filter membrane and using probe hydrization for fragment detection.

Southern Blotting is used to determine whether a DNA sample contains a specific sequence or if it does not, and to see the size of the restriction enzyme that contains the sequence. It is also used to test for certain mutations in the DNA that is commonly associated with genetic diseases. Southern Blotting is also used for identification. Scientists build a genetic profile to identify certain sequences. (I.E.: forensics and paternity testing)

1. Use a restriction enzyme to digest or fragment the DNA sequence. 2. Run the sample fragment on an agarose gel. 3. While the DNA is still on the gel, depurinate and denature the DNA using an HCL (depurination) and NaCl solution. 4. Transfer the gel containing the denatured DNA to the membrane wells down.. The membrane may be nitrocellulose or nylon. Nylon is more commonly used due to it being easier to bind to and less fragile compared to a nitrocellulose membrane. 5. Blotting filter will be placed on top of the membrane, paper towels stacked on top, and then a weight. 6. After some time passes, a blue ink pen is used to mark the wells positions on the membrane before disposing of the gel.

6. Using an oven, the membrane is dried through baking. Or it can simply be dried with filter paper. 7. Once dry, the membrane must be put on a blot solution. The dye must then be washed away and the bands of DNA should be visible. 8. A probe may be used to visualize a certain/targeted sequence. The probe can be seen through X-Ray.