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Mismatch discrimination in oligonucleotide hybridization reactions using single strand binding protein. A surface plasmon fluorescence study
Author(s) -
Neumann Thomas,
Knoll Wolfgang
Publication year - 2001
Publication title -
israel journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.908
H-Index - 54
eISSN - 1869-5868
pISSN - 0021-2148
DOI - 10.1560/f0q3-whxd-t7vh-jj78
Subject(s) - chemistry , fluorophore , oligonucleotide , fluorescence , dissociation constant , surface plasmon , surface plasmon resonance , fluorescence spectroscopy , biophysics , photochemistry , plasmon , dna , biochemistry , nanotechnology , nanoparticle , receptor , physics , materials science , quantum mechanics , biology
We describe the use of surface plasmon‐ and surface plasmon fieldenhanced fluorescence spectroscopy for the detection of hybridization reactions between surface‐attached probe oligonucleotides and complement strands binding from solution. These targets, exhibiting different base mismatches relative to the probe 15‐mer sequences, carry a fluorophore at their 5'‐end thus allowing for sensitive detection and quantification of association, k on , and dissociation, k off , rate constants, as well as affinity constants, K a . We demonstrate that by the competitive binding / replacement of single strand binding proteins the mismatch discrimination can be further enhanced.