Astrategy for rapid and efficient DNA sequencing by mass spectrometry

Hubert Köster, Kai Tang, Dong Jing Fu, Andreas Braun, Dirk Van Den Boom, Cassandra L. Smith, Robert J. Cotter, Charles R. Cantor

Research output: Contribution to journalArticle

Abstract

Two methods of solid-phase Sanger DNA sequencing followed by detection with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry are demonstrated. In one method, sequencing ladders generated on an immobilized synthetic template were resolved up to the 63-mer including the primer. Detection sensivity and resolution were sufficient for sequence analysis in the given range. This approach is particularly suitable for comparative (diagnostic) DNA sequencing. A second method that has the potential for high throughput de novo DNA sequencing is also presented; it uses immobilized duplex probes with five-base single-stranded overhangs to capture an unknown DNA template serving as primers for Sanger DNA sequencing. The power of mass spectrometry is demonstrated not only by its very high speed, but also by its ability to identify sequences that are not readable using gel electrophoresis.

Original languageEnglish (US)
Pages (from-to)1123-1128
Number of pages6
JournalNature Biotechnology
Volume14
Issue number9
StatePublished - Sep 1996

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DNA Sequence Analysis
Mass spectrometry
Mass Spectrometry
DNA
DNA Primers
Ladders
Electrophoresis
Ionization
Sequence Analysis
Desorption
Lasers
Gels
Throughput

Keywords

  • DNA sequencing
  • mass spectrometry
  • solid phase

ASJC Scopus subject areas

  • Microbiology

Cite this

Köster, H., Tang, K., Fu, D. J., Braun, A., Van Den Boom, D., Smith, C. L., ... Cantor, C. R. (1996). Astrategy for rapid and efficient DNA sequencing by mass spectrometry. Nature Biotechnology, 14(9), 1123-1128.

Astrategy for rapid and efficient DNA sequencing by mass spectrometry. / Köster, Hubert; Tang, Kai; Fu, Dong Jing; Braun, Andreas; Van Den Boom, Dirk; Smith, Cassandra L.; Cotter, Robert J.; Cantor, Charles R.

In: Nature Biotechnology, Vol. 14, No. 9, 09.1996, p. 1123-1128.

Research output: Contribution to journalArticle

Köster, H, Tang, K, Fu, DJ, Braun, A, Van Den Boom, D, Smith, CL, Cotter, RJ & Cantor, CR 1996, 'Astrategy for rapid and efficient DNA sequencing by mass spectrometry', Nature Biotechnology, vol. 14, no. 9, pp. 1123-1128.
Köster H, Tang K, Fu DJ, Braun A, Van Den Boom D, Smith CL et al. Astrategy for rapid and efficient DNA sequencing by mass spectrometry. Nature Biotechnology. 1996 Sep;14(9):1123-1128.
Köster, Hubert ; Tang, Kai ; Fu, Dong Jing ; Braun, Andreas ; Van Den Boom, Dirk ; Smith, Cassandra L. ; Cotter, Robert J. ; Cantor, Charles R. / Astrategy for rapid and efficient DNA sequencing by mass spectrometry. In: Nature Biotechnology. 1996 ; Vol. 14, No. 9. pp. 1123-1128.
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