Ratiometric FRET-based detection of DNA and micro-RNA on the surface using TIRF detection

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

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

A new FRET-based method for the ratiometric detection of DNA oligomers on a surface using TIRF detection mode is reported. The dual-labeled system consisting of two hybridized oligomers, Cy3oligoY:Cy5oligoX was immobilized on the surface, and the total internal reflection fluorescence (TIRF) was used to detect emission signals from the surface. Two signals, green and red, which originated from the green donor Cy3 and the red acceptor Cy5, have been simultaneously detected. When the target single-stranded complimentary oligomer was present in the solution, this oligomer replaced the Cy3oligoY in the donor:acceptor complex on the surface and the ratio of red-to-green signal was dramatically changed. This detection scheme is generally applicable to the detection of DNA or RNA on a surface.

Original languageEnglish
Pages (from-to)698-702
Number of pages5
JournalJournal of Luminescence
Volume130
Issue number4
DOIs
StatePublished - 1 Apr 2010

Keywords

  • DNA and RNA detection
  • DNA hybridization
  • FRET
  • Förster resonance energy transfer
  • Ratiometric fluorescence assay
  • Total internal reflection fluorescence

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