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

doi:10.1016/j.jlumin.2009.11.022

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 journal › Article › peer-review

9 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 language	English
Pages (from-to)	698-702
Number of pages	5
Journal	Journal of Luminescence
Volume	130
Issue number	4
DOIs	https://doi.org/10.1016/j.jlumin.2009.11.022
State	Published - 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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10.1016/j.jlumin.2009.11.022

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@article{728528c0f7d64ed6a7c40c863a6166a6,

title = "Ratiometric FRET-based detection of DNA and micro-RNA on the surface using TIRF detection",

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.",

keywords = "DNA and RNA detection, DNA hybridization, FRET, F{\"o}rster resonance energy transfer, Ratiometric fluorescence assay, Total internal reflection fluorescence",

author = "Matveeva, {Evgenia G.} and Zygmunt Gryczynski and Stewart, {Donald R.} and Ignacy Gryczynski",

note = "Funding Information: This work was supported by the NIH R41EB008614-01A1 grant (D.S.), Texas Emerging Technology Fund grant (Z.G.), and ARP ATP Grant 000130-0042-2007 (I.G.).",

year = "2010",

month = apr,

doi = "10.1016/j.jlumin.2009.11.022",

language = "English",

volume = "130",

pages = "698--702",

journal = "Journal of Luminescence",

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publisher = "Elsevier",

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T1 - Ratiometric FRET-based detection of DNA and micro-RNA on the surface using TIRF detection

AU - Matveeva, Evgenia G.

AU - Gryczynski, Zygmunt

AU - Stewart, Donald R.

AU - Gryczynski, Ignacy

N1 - Funding Information: This work was supported by the NIH R41EB008614-01A1 grant (D.S.), Texas Emerging Technology Fund grant (Z.G.), and ARP ATP Grant 000130-0042-2007 (I.G.).

PY - 2010/4

Y1 - 2010/4

N2 - 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.

AB - 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.

KW - DNA and RNA detection

KW - DNA hybridization

KW - FRET

KW - Förster resonance energy transfer

KW - Ratiometric fluorescence assay

KW - Total internal reflection fluorescence

UR - http://www.scopus.com/inward/record.url?scp=74349117811&partnerID=8YFLogxK

U2 - 10.1016/j.jlumin.2009.11.022

DO - 10.1016/j.jlumin.2009.11.022

M3 - Article

AN - SCOPUS:74349117811

SN - 0022-2313

VL - 130

SP - 698

EP - 702

JO - Journal of Luminescence

JF - Journal of Luminescence

IS - 4

ER -

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

Abstract

Keywords

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