Ratiometric FRET-based detection of DNA and micro-RNA in solution

Evgenia G. Matveeva, Zygmunt Gryczynski, Donald R. Stewart, Ignacy Gryczynski

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

A ratiometric method for detecting DNA oligomers in bulk solution based on Förster resonance energy transfer (FRET) is described. The two fluorescence signals (green and red), originating from Cy3 (donor, green) and Cy5 (acceptor, red) labels, are simultaneously detected from the pre-hybridized Cy3oligomerY:Cy5oligomerX system. The ratio of red to green intensities is sensitive to the presence of the single-stranded complimentary oligomer, which replaces single-stranded Cy3oligomerY in the donor:acceptor complex and perturbs the FRET. The detection scheme is generally applicable to the detection of DNA and RNA, and particularly micro-RNA. The proposed method is applicable to various double-stranded various lengths targets (manipulation of the sample preparation conditions, such as temperature, incubation time, denaturizing agent, may be needed).

Original languageEnglish
Pages (from-to)1281-1285
Number of pages5
JournalJournal of Luminescence
Volume129
Issue number11
DOIs
StatePublished - 1 Nov 2009

Keywords

  • DNA and micro-RNA detection
  • DNA hybridization
  • Förster resonance energy transfer
  • Ratiometric fluorescence assay

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