Jean-Claude Bunzli - A versatile method for quantification of DNA and PCR products based on time-resolved Eu(III) luminescence

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      Publication Details (including relevant citation   information):

      Song,B., Vandevyver,C.D.B., Deiters,E., Chauvin,A.S., Hemmila,I.,   Bunzli,J.C.G. Analyst 2008 133  (12) 1749-1756

      Abstract: A versatile and robust method for the   determination of DNA and PCR products (<500 bp) is presented,   based on a mix of an Eu(III) chelate and acridine orange (AO).   The nucleic acid selective stains acridine orange (AO) and   ethidium bromide (EB) quench the luminescence of the bimetallic   [Eu(2)(L(C2))(3)] and of other monometallic chelates such as the   macrocyclic complex [Eu(L(kel))], even at very low molar ratios.   Stern-Volmer plots of the metal-centered emission intensities   (F(0)/F) and Eu((5)D(0)) lifetimes (tau(0)/tau) show the AO   quenching being purely dynamic with K(D) 6.7 x 10(5) M(-1) for   [Eu(2)(L(C2))(3)] and 1.6 x 10(6) M(-1) for [Eu(L(kel))], and   bimolecular rate constants k(q) 2.7 x 10(8) M(-1) s(-1) and 3.4 x   10(9) M(-1) s(-1), respectively. On the other hand, EB quenching   is due to both dynamic and static mechanisms. In the presence of   various types of DNA > 0.1 ng mu L(-1) (dsDNA, ssDNA or   circular DNA), the quenched luminescence is reinstated, AO and EB   intercalating into DNA, which removes the interaction with the   Eu(III) complexes. The best results are obtained with   [Eu(2)(L(C2))(3)]/AO with detections limits in the range   0.18-0.66 ng mu L(-1); detection limits for the [Eu(L(kel))]/AO   system are slightly larger; simpler monometallic Eu(III)   complexes with dipicolinate derivatives do not follow suit in   that they decompose in the presence of DNA. The Eu(III)/AO method   is shown to be pH insensitive in the range 3-10; furthermore it   is essentially insensitive to 1000-fold excesses of potential   interfering substances, e. g. BSA, glucose, chelating agents and   anions, alkaline earth and transition metal cations, variations   in luminescence intensity being < 5%, (10 analytes) or 5-10%   (4 analytes); only Co(II) and Cu(II) interfere substantially

      Address (URL): WOS:000261005100015