Time-resolved fluorimetric detection of terbium-labelled deoxyribonucleic acid separated by gel electrophoresis

Steven S Saavedra, Enrico G. Picozza

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Deoxyribonucleic acid (DNA) was reacted with a strong chelating agent and labelled with terbium, yielding a highly fluorescent conjugate with a lifetime of 1.5 ms. When a pulsed source and gated detection electronics were employed, the long-lived decay allowed effective discrimination against background fluorescence and scattered excitation. Detection limits should therefore be signifcantly improved in comparison with covalent labels with fluorescence lifetimes in the nanosecond regime or stains such as ethidium bromide. The conjugate is very stable, remaining fluorescent on dilution and in an electric field at elevated temperatures (60 °C), conditions typically encountered during polyacrylamide gel electrophoresis. As an enhancement solution is not required to develop the fluorescence, this system could be utilised in situations where on-line detection is desirable. Although only DNA was labelled, the method is equally applicable to ribonucleic acid.

Original languageEnglish (US)
Pages (from-to)835-838
Number of pages4
JournalAnalyst
Volume114
Publication statusPublished - 1989
Externally publishedYes

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Keywords

  • Deoxyribonucleic acid detection
  • Gel electrophoresis
  • Nucleic acids
  • Terbium chelate
  • Time-resolved fluorescence

ASJC Scopus subject areas

  • Analytical Chemistry

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