TY - JOUR
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 -