SOUTHERN BLOTTING
Principle: ▪ Southern blotting is an example of RFLP (restriction fragment length polymorphism). It was developed by Edward M. Southern (1975). Southern blotting is a hybridization technique for identification of particular size of DNA from the mixture of other similar molecules. This technique is based on the principle of separation of DNA fragments by gel electrophoresis and identified by labelled probe hybridization. ▪ Basically, the DNA fragments are separated on the basis of size and charge during electrophoresis. Separated DNA fragments after transferring on nylon membrane, the desired DNA is detected using specific DNA probe that is complementary to the desired DNA. ▪ A hybridization probe is a short (100-500bp), single stranded DNA. The probes are labeled with a marker so that they can be detected after hybridization. Procedure/ Steps: 1. Restriction digest: by RE enzyme and amplification by PCR 2. Gel electrophoresis: SDS gel electrophoresis 3. Denaturation: Treating wi
