Simultaneous On-line DNA Sequencing on Both Strands with Two Fluorescent Dyes

S. Wiemann, J. Stegemann, D. Grothues, A. Bosch, Xavier P. Estivill, C. Schwager, J. Zimmermann, H. Voss, W. Ansorge

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34 Citations (Scopus)

Abstract

We describe an automated DNA-sequencing technique which allows both the simultaneous sequencing from the two strands of double-stranded templates and the subsequent detection of the sequencing products on-line and in parallel. The technique is based on hardware technology also used in the ALF DNA sequencer (Pharmacia, Uppsala). A helium-neon laser was mounted into the sequencing device additionally to the standard argon laser. Two different primers, labeled with either fluorescein or Texas red, are used in a single sequencing reaction resulting in an output of two sequences. Both sequencing products are then analyzed on-line in the same lanes of a gel. This technique is especially useful for the complete sequencing of DNA fragments up to 1 kb. High accuracy sequencing of PCR products in double-stranded form can now be accomplished in only one sequencing reaction.

Original languageEnglish
Pages (from-to)117-121
Number of pages5
JournalAnalytical Biochemistry
Volume224
Issue number1
DOIs
Publication statusPublished - Jan 1995
Externally publishedYes

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wiemann, S., Stegemann, J., Grothues, D., Bosch, A., Estivill, X. P., Schwager, C., Zimmermann, J., Voss, H., & Ansorge, W. (1995). Simultaneous On-line DNA Sequencing on Both Strands with Two Fluorescent Dyes. Analytical Biochemistry, 224(1), 117-121. https://doi.org/10.1006/abio.1995.1015