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

Research output: Contribution to journalArticle

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

Fingerprint

DNA Sequence Analysis
Fluorescent Dyes
Neon
Gas Lasers
Argon
DNA
Fluorescein
Helium neon lasers
Argon lasers
Lasers
Gels
Technology
Equipment and Supplies
Polymerase Chain Reaction
Hardware
Texas red

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., ... 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

Simultaneous On-line DNA Sequencing on Both Strands with Two Fluorescent Dyes. / Wiemann, S.; Stegemann, J.; Grothues, D.; Bosch, A.; Estivill, Xavier P.; Schwager, C.; Zimmermann, J.; Voss, H.; Ansorge, W.

In: Analytical Biochemistry, Vol. 224, No. 1, 01.1995, p. 117-121.

Research output: Contribution to journalArticle

Wiemann, S, Stegemann, J, Grothues, D, Bosch, A, Estivill, XP, Schwager, C, Zimmermann, J, Voss, H & Ansorge, W 1995, 'Simultaneous On-line DNA Sequencing on Both Strands with Two Fluorescent Dyes', Analytical Biochemistry, vol. 224, no. 1, pp. 117-121. https://doi.org/10.1006/abio.1995.1015
Wiemann, S. ; Stegemann, J. ; Grothues, D. ; Bosch, A. ; Estivill, Xavier P. ; Schwager, C. ; Zimmermann, J. ; Voss, H. ; Ansorge, W. / Simultaneous On-line DNA Sequencing on Both Strands with Two Fluorescent Dyes. In: Analytical Biochemistry. 1995 ; Vol. 224, No. 1. pp. 117-121.
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