DNA sequence analysis is one of the pivotal methods of modern molecular biology. It plays a central role in virtually every project that involves the cloning, characterization, and manipulation of RNA or DNA. Since the development of rapid sequencing techniques in the early 1970s, more than 20 million bases have been sequenced by manual techniques.
This DNA sequencing has all been accomplished with either of two techniques: the enzymatic method of sequencing, developed by Sanger and Coulson, and the chemical method, developed by Maxam and Gilbert. In both methods, as they are conventionally practiced, sets of radiolabeled DNA fragments with a common origin, but terminating at a particular base or bases, are produced in the sequencmg reactions. These fragment sets are loaded in adjacent lanes of a high-resolution polyacrylamide slab gel separated by electrophoresis, and a film image is obtained by autoradiography. Analysis of the autoradiogram yields the DNA sequence ...
