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Nature 321, 674-679 (12 June 1986) | doi:10.1038/321674a0; Received 30 December 1985; Accepted 11 April 1986

Fluorescence detection in automated DNA sequence analysis

Lloyd M. Smith, Jane Z. Sanders, Robert J. Kaiser, Peter Hughes, Chris Dodd, Charles R. Connell*, Cheryl Heiner*, Stephen B. H. Kent & Leroy E. Hood

  1. Division of Biology, California Institute of Technology, Pasadena, California 91125, USA
  2. *Applied Biosystems, Inc., Foster City, California 94404, USA
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We have developed a method for the partial automation of DNA sequence analysis. Fluorescence detection of the DNA fragments is accomplished by means of a fluorophore covalently attached to the oligonucleotide primer used in enzymatic DNA sequence analysis. A different coloured fluorophore is used for each of the reactions specific for the bases A, C, G and T. The reaction mixtures are combined and co-electrophoresed down a single polyacrylamide gel tube, the separated fluorescent bands of DNA are detected near the bottom of the tube, and the sequence information is acquired directly by computer.