Chapter 6 outlines the range of methods used to isolate, purify and analyse nucleic acids. Methods to quantify DNA and RNA, and labelling of nucleic acids using radioactive and fluorographic precursors and a range of enzymatic methods, are described. The use of gel electrophoresis to separate DNA fragments is discussed. The principles of first-generation DNA sequencing are outlined, and the Sanger dideoxy method described for manual and automated methods. Next-generation methods for DNA sequencing are covered, to illustrate the range of advanced techniques that have enabled large-scale genome sequencing to become a routine laboratory procedure that is both rapid and cost-effective. Techniques for massively parallel and single-molecule real-time sequencing are described.
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