Determination of binding affinities of triplex forming oligonucleotides using a fluorescent intercalator displacement (FID) assay
The binding affinities of several triplex forming oligonucleotides were determined using a fluorescent intercalator displacement (FID) assay.
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Results and Discussion
Below, we report the binding affinities of several triplex forming oligonucleotides with homopurine homopyrimidine hairpins consisting of either a pure T·AT and C+·GC triad (hairpins 1 and 2, 9-mer TFOs) or incorporating both T·AT and C+·GC triads (hairpins 3 and 4, 13-mer TFOs) (Fig. 3).
The results are summarized in Table 1. In each case of triplex formation, a well defined titration curve was observed exhibiting the expected 1:1 stoichiometry of binding and providing association constants
Acknowledgements
We gratefully acknowledge the financial support of NIH (CA 41986, CA 78045) and the Skaggs Institute for Chemical Biology. W.C.T. is a Skaggs Fellow and recipient of an ACS Medicinal Chemistry Division Graduate Fellowship sponsored by Wyeth Ayerst (2002–2003).
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