Colorimetric allele analysis based on the DNA-directed cooperative formation of luminous lanthanide complexes.
We present the facile technique of colorimetric SNP analysis through DNA-templated cooperative complexation between a luminescent lanthanide ion (Ln(3+): Tb(3+) or Eu(3+)) 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 Ln(3+). The capture probes for the wild-type (wt) and the mutant (mut) of thiopurine S-methyltransferase gene, were mixed with equimolar amounts of Tb(3+) and Eu(3+), 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.
Kitamura
Yusuke
Y
Department of Applied Chemistry and Biochemistry, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan.
Ihara
Toshihiro
T
Tsujimura
Yusuke
Y
Osawa
Yuka
Y
Jyo
Akinori
A
D000483:Alleles; D002614:Chelating Agents / Q000737:chemistry; D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000737:chemistry; D004492:Edetic Acid / Q000737:chemistry; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D049408:Luminescent Agents / Q000737:chemistry*; D008780:Methyltransferases / Q000235:genetics; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D015345:Oligonucleotide Probes / Q000737:chemistry*; D010618:Phenanthrolines / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide; D013053:Spectrophotometry
D002614:Chelating Agents; D028581:Lanthanoid Series Elements; D049408:Luminescent Agents; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D010618:Phenanthrolines; D004247:DNA; D004492:Edetic Acid; D008780:Methyltransferases; C022745:thiopurine methyltransferase; C025205:1,10-phenanthroline