Mismatch discrimination in oligonucleotide hybridization reactions using single strand binding protein. A surface plasmon fluorescence study |
| |
Authors: | Thomas Neumann Wolfgang Knoll |
| |
Affiliation: | Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany |
| |
Abstract: | 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, kon, and dissociation, koff, rate constants, as well as affinity constants, Ka. We demonstrate that by the competitive binding / replacement of single strand binding proteins the mismatch discrimination can be further enhanced. |
| |
Keywords: | |
|
|