Abstract

DNA sequencing is a necessary technique for almost all molecular biology studies including DNA cloning, characterization, mutagenesis, DNA recombination, regulation of gene expression, ¹ – ³ and protein expression. There are several well-established methods for nucleic acid sequencing, and in recent years, advances in automated sequencing have made DNA sequence analysis an easy and routine process. The present chapter describes in detail the preparation of DNA for sequencing. ⁴ – ⁶ DNA sequencing by dideoxynucleotides chain termination^2,3 and cycle sequencing. ¹ , ⁴ – ⁶ These techniques are the foundation of most sequencing applications; however, with the advent of automated sequencing and the availability of automated liquid handlers, we also briefly describe how the newer machines and sources have greatly increased the speed of DNA sequence analysis. The following techniques are routinely used in our respective labs.