Journal
JOURNAL OF LUMINESCENCE
Volume 129, Issue 11, Pages 1281-1285Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jlumin.2009.06.009
Keywords
Ratiometric fluorescence assay; Forster resonance energy transfer; DNA hybridization; DNA and micro-RNA detection
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Funding
- BITC
- ARP ATP [000130-0042-2007]
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A ratiometric method for detecting DNA oligomers in bulk solution based on Forster 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). (c) 2009 Elsevier B.V. All rights reserved.
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