Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Real Time Automated Simultaneous Double-Stranded DNA Sequencing Using Two-Color Fluorophore Labeling

Abstract

We determined the base sequences of both strands of duplex DNA simultaneously with an automated DNA sequencer. Primers for each strand were labeled with two different dyes and chain extension reactions for both strands produced four DNA families: “A”, “C”, “G”, and “T”. An image splitting-prism and band-pass filters were mounted on a real time DNA sequencer to detect fluorescences from the two different dyes separately. As a result, one thousand bases of a clone could be determined at the same time.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Smith, L.M., Sanders, J.Z., Kaiser, R.J., Hughes, P., Dodd, C., Connell, C.R., Heiner, C., Kent, S.B.H. and Hood, L.E. 1986. Fluorescence detection in automated DNA sequence analysis. Nature 321: 674–679.

    Article  CAS  Google Scholar 

  2. Ansorge, W., Sproat, B.S., Stegemann, J. and Schwager, C. 1986. A non-radio active automated method for DNA sequencing determination. J. Biochem. Biophys. Methods 13: 315–323.

    Article  CAS  Google Scholar 

  3. Prober, J.M., Trainor, G.L., Dam, R.J., Hobbs, F.W., Robertson, C.W., Zagursky, R.J., Cocuzza, A.J., Jensen, M.A. and Baumeister, K. 1987. A system for rapid DNA sequencing with fluorescent chain-terminating dideoxy nucleotides. Science 238: 336–341.

    Article  CAS  Google Scholar 

  4. Kambara, H., Nishikawa, T., Katayama, Y. and Yamaguchi, T. 1988. Optimization of parameters in a DNA sequenator using fluorescence detection. Bio/Technology 6: 816–821.

    CAS  Google Scholar 

  5. Luckey, J.A., Drossman, H., Kostichka, A.J., Mead, D.A., D'Cunha, J., Norris, T.B. and Smith, L.M. 1990. High speed DNA sequencing by capillary electrophoresis. Nuc. Acids Res. 18: 4417–4421.

    Article  CAS  Google Scholar 

  6. Tabor, S. and Richardson, C.C. 1989. Effect of manganese ions on the incorporations of dideoxy nucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase I. Proc. Natl. Acad. Sci. USA 86: 4076–4080.

    Article  CAS  Google Scholar 

  7. Zimmermann, J., Voss, H., Schwager, C., Stegemann, J., Erfle, H., Stucky, K., Kistensen, T. and Ansorge, W. 1990. A simplified protocol for fast plasmid DNA sequencing. Nuc. Acids Res. 18: 1067.

    Article  CAS  Google Scholar 

  8. Church, G.M. and Kieffer-Higgins, S. 1988. Multiplex DNA sequencing. Science 240: 185–188.

    Article  CAS  Google Scholar 

  9. Yang, M.M. and Youvan, D.C. 1989. A prospectus for multispectral-multiplex DNA sequencing. Bio/Technology 7: 576–580.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kambara, H., Nagai, K. & Hayasaka, S. Real Time Automated Simultaneous Double-Stranded DNA Sequencing Using Two-Color Fluorophore Labeling. Nat Biotechnol 9, 648–651 (1991). https://doi.org/10.1038/nbt0791-648

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0791-648

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing