Top : Molecular Biology : DNA : Southern Blotting

Summary: Southern blotting was named after Edward M. Southern who developed this procedure at Edinburgh University in the 1975. It allows investigators to determine the molecular weight of a restriction fragment, to measure relative amounts in different samples and to locate a particular sequence of DNA within a complex mixture. DNA (genomic or other source) is digested with a restriction enzyme and separated by gel electrophoresis and transferred from an agarose gel onto a membrane which is then incubated with a probe which is single-stranded DNA. This probe will form base pairs with its complementary DNA sequence and bind to form a double-stranded DNA molecule. The probe is labeled before hybridization either radioactively or enzymatically (e.g. alkaline phosphatase or horseradish peroxidase). Finally, the location of the probe is detected by directly exposing the membrane to X-ray film or chemiluminescent methods.


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