Colorimetric allele analysis based on the DNA-directed cooperative formation of luminous lanthanide complexes - DNA

Colorimetric allele analysis based on the DNA-directed cooperative formation of luminous lanthanide complexes
2006
Yusuke Kitamura1, Toshihiro Ihara1,2, Yusuke Tsujimura1, Yuka Osawa1 and Akinori Jyo1
1 Department of Applied Chemistry and Biochemistry, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan, 2 PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho Kawaguchi, Saitama 332-0012, Japan
Nucleic Acids Symposium Series 2006
We present the facile technique of colorimetric SNP analysis through DNA-templated cooperative complexation between a luminescent lanthanide ion (Ln3+: Tb3+ or Eu3+) and two ODN (oligodeoxyribonucleotide) conjugates carrying a metal chelator. Ethylenediaminetetraacetic acid (EDTA) and 1,10-phenanthrorine (phen) were covalently attached to ODNs to form the conjugate probes, capture and sensitizer probes, respectively. The sequences of the conjugates were designed so as to form a tandem duplex with a target with their auxiliary units facing each other, providing a microenvironment to accommodate Ln3+. The capture probes for the wild-type (wt) and the mutant (mut) of thiopurine S-methyltransferase gene, were mixed with equimolar amounts of Tb3+ and Eu3+, respectively. Then both of the allele specific capture probe solutions and the sensitizer probe were added to three different solutions containing the targets, wt/wt (G/G), mut/mut (C/C), and wt/mut (G/C). The solutions emitted in green, red, and yellow, respectively; the colors were identified even by the naked eye.
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