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Fluorescence detection in automated DNA sequence analysis

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Abstract

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.

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Author notes

  1. Charles R. Connell and Cheryl Heiner: Applied Biosystems, Inc., Foster City, California 94404, USA

Authors and Affiliations

  1. Division of Biology, California Institute of Technology, Pasadena, California, 91125, USA

    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

Authors
  1. Lloyd M. Smith
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  2. Jane Z. Sanders
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  3. Robert J. Kaiser
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  4. Peter Hughes
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  5. Chris Dodd
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  6. Charles R. Connell
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  7. Cheryl Heiner
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  8. Stephen B. H. Kent
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  9. Leroy E. Hood
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Smith, L., Sanders, J., Kaiser, R. et al. Fluorescence detection in automated DNA sequence analysis. Nature 321, 674–679 (1986). https://doi.org/10.1038/321674a0

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  • DOI: https://doi.org/10.1038/321674a0

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