TY - JOUR
T1 - Single-molecule Fluorescence Spectroscopy of TOTO on Poly-AT and Poly-GC DNA
AU - Bowen, Benjamin P.
AU - Enderlein, Jörg
AU - Woodbury, Neal
PY - 2003/12
Y1 - 2003/12
N2 - Excited state lifetime and amplitude measurements were made on thiazole orange dimer (TOTO), a dimeric DNA-intercalating fluorophore, at single-molecule concentrations. As expected from previous study, the excited state lifetime of TOTO intercalated in DNA is dependent on the sequence of the double-stranded DNA, having values of 2.2 ns in poly-GC DNA and 1.8 ns in poly-AT DNA. The distribution of excited state lifetimes of single molecules of TOTO intercalated into oligonucleotides having varying proportions of poly-GC sequences relative to poly-AT sequences were analyzed as a function of the fraction of poly-GC. By using excited state lifetime distributions from the purely GC and purely AT oligonucleotides as a basis set, it was possible to estimate the GC content of oligonucleotides with intermediate GC composition to within a few percent error. This serves as a model for the analysis of equilibrium binding distributions in DNA using single-molecule methods.
AB - Excited state lifetime and amplitude measurements were made on thiazole orange dimer (TOTO), a dimeric DNA-intercalating fluorophore, at single-molecule concentrations. As expected from previous study, the excited state lifetime of TOTO intercalated in DNA is dependent on the sequence of the double-stranded DNA, having values of 2.2 ns in poly-GC DNA and 1.8 ns in poly-AT DNA. The distribution of excited state lifetimes of single molecules of TOTO intercalated into oligonucleotides having varying proportions of poly-GC sequences relative to poly-AT sequences were analyzed as a function of the fraction of poly-GC. By using excited state lifetime distributions from the purely GC and purely AT oligonucleotides as a basis set, it was possible to estimate the GC content of oligonucleotides with intermediate GC composition to within a few percent error. This serves as a model for the analysis of equilibrium binding distributions in DNA using single-molecule methods.
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U2 - 10.1562/0031-8655(2003)078<0576:SFSOTO>2.0.CO;2
DO - 10.1562/0031-8655(2003)078<0576:SFSOTO>2.0.CO;2
M3 - Article
C2 - 14743865
AN - SCOPUS:0345743716
SN - 0031-8655
VL - 78
SP - 576
EP - 581
JO - Photochemistry and photobiology
JF - Photochemistry and photobiology
IS - 6
ER -