TY - JOUR
T1 - Ratiometric FRET-based detection of DNA and micro-RNA in solution
AU - Matveeva, Evgenia G.
AU - Gryczynski, Zygmunt
AU - Stewart, Donald R.
AU - Gryczynski, Ignacy
N1 - Funding Information:
This work was supported by the Texas Emerging Technologies Fund grant, BITC, and ARP ATP Grant 000130-0042-2007 (I.G.). Authors thank Maria Matveeva for the help with figures drawing.
PY - 2009/11
Y1 - 2009/11
N2 - 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).
AB - 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).
KW - DNA and micro-RNA detection
KW - DNA hybridization
KW - Förster resonance energy transfer
KW - Ratiometric fluorescence assay
UR - http://www.scopus.com/inward/record.url?scp=68949162045&partnerID=8YFLogxK
U2 - 10.1016/j.jlumin.2009.06.009
DO - 10.1016/j.jlumin.2009.06.009
M3 - Article
AN - SCOPUS:68949162045
SN - 0022-2313
VL - 129
SP - 1281
EP - 1285
JO - Journal of Luminescence
JF - Journal of Luminescence
IS - 11
ER -