Demystified ... DNA nucleotide sequencing.

The determination of the nucleotide sequence of a gene is often the ultimate goal of a research project and the frequency with which new sequences are described in the literature and computer databases is remarkable. The basic techniques that underpin these endeavours are remarkably simple and their description now frequently forms part of the very earliest portion of a biological science undergraduate degree. Despite this, many misunderstandings about the nature of the reactions occur. In this review, we consider the mechanics of the techniques, including aspects of the associated technologies such as polyacrylamide gel electrophoresis. We describe the type of DNA suitable for sequencing and how it may be prepared, the details of the enzymatic reactions that can be used by the researcher, and the modifications to the techniques that have allowed the development of automation, which has made the goal of determining the nucleotide sequence of the entire human genome a reality.