Detection of single base changes in nucleic acids.

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Detection of single base changes in nucleic acids.

Publication Date
Oct 01, 1989
  • Biology
  • Medicine


Biochem. J. (1989) 263, 1-10 (Printed in Great Britain) REVIEW ARTICLE Detection of single base changes in nucleic acids Richard G. H. COTTON Murdoch Institute, Royal Children's Hospital, Melbourne, Victoria 3052, Australia INTRODUCTION Ever since the detection of the first mutations in Escherichia coli, and subsequently in human DNA (Flavel et al., 1978), enormous efforts have been expended in defining changes in nucleic acid sequences of many organisms. This activity has been pursued in order to: (a) understand mechanisms of disease, (b) understand the structure and function of enzymes and nucleic acids (e.g. tRNA), (c) track micro-organisms in the environment, (d) track disease genes in people for diagnostic purposes, (e) define mutagenic substances, and (f) discover evol- utionary relationships between species. Faced with the tedium and repetitiveness of cloning and sequencing a gene containing a mutation which may take 6 months to define, there has been increasing interest in shortening this process and the last decade has seen some of the fruits of this activity. It has proven difficult to develop methods which will simply detect all or most point mutation types (see below) and hence all or most mutations in a given stretch of nucleic acid. The end point of these studies should be a method which will guarantee detection of all point mutations with a mini- mum of manipulation of the genetic material before application of a method of extreme simplicity. This review traces the history and evolution of meth- odology which has increased the rate of definition of single base mutations. It is not intended as a review of current methods for studying human disease as this is covered elsewhere (Landegren et al., 1988b; Caskey, 1987). Particular methods have been reviewed recently (Myers et al., 1988; Myers & Maniatis, 1986; Lerman et al., 1984). The techniques covered fall into three broad categories: (a) methods for cloning (amplification) and sequencing, (b) methods using hybrid molecules (het


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