Single-molecule interactions studied by using a modified DNA sequencer: A comparison with surface plasmon resonance data

In this study we established the principles for using a commercial off-the-shelf DNA sequencer (RS, Pacific Biosciences) to measure biomolecular interactions. Binding between single oligonucleotides immobilized at the bottom of a nanowell to a complementary DNA strand was used as a model system. The influence of the labeling dye on the hybridization of a 7mer oligonucleotide was investigated. The resulting association and dissociation rate constants (kon and koff) and the calculated dissociation constants (Kd) were compared to data obtained from surface plasmon resonance (SPR) measurements. In the present study we identified a good agreement of the determined kinetic constants by the two methods investigated.