Determination of nucleotide sequence in DNA is the important technique in molecular biology. This technique was first developed by the two scientist groups in 1975. Maxam AM and Gilbert W. American scientists at Havard University developed the nucleotide sequencing technique using the chemical method. Sanger F, Niklen S and Coulson AR, the Bristish scientists at Cambridge successfully developed this technique using the enzymatic method. Nowadays these techniques are widely used in molecular biology research. The strategies for DNA sequencing consists of 4 steps: 1. Preparation of DNA template 2. Sequencing reaction 3. Electrophoresis 4. Analysis 1. Preparation of DNA template The DNA template used for sequencing can be prepared by several methods: a) Gene cloning It is conventional method. Isolate and purify genomic DNA. Cut with appropriate restriction enzyme. Cut sequencing vector with the same enzyme. Insert DNA ...
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