Specific Enzymatic Amplification of DNA In Vitro: The Polymerase Chain Reaction

  1. K. Mullis,
  2. F. Faloona,
  3. S. Scharf,
  4. R. Saiki,
  5. G. Horn, and
  6. H. Erlich
  1. Cetus Corporation, Department of Human Genetics, Emeryville, California 94608

This extract was created in the absence of an abstract.

Excerpt

The discovery of specific restriction endonucleases (Smith and Wilcox 1970) made possible the isolation of discrete molecular fragments of naturally occurring DNA for the first time. This capability was crucial to the development of molecular cloning (Cohen et al. 1973); and the combination of molecular cloning and endonuclease restriction allowed the synthesis and isolation of any naturally occurring DNA sequence that could be cloned into a useful vector and, on the basis of flanking restriction sites, excised from it. The availability of a large variety of restriction enzymes (Roberts 1985) has significantly extended the utility of these methods.

The de novo organic synthesis of oligonucleotides and the development of methods for their assembly into long double-stranded DNA molecules (Davies and Gassen 1983) have removed, at least theoretically, the minor limitations imposed by the availability of natural sequences with fortuitously unique flanking restriction sites. However, de novo synthesis, even with automated...

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