Electrokinetic Method For Single Molecule DNA Sequencing

DESCRIPTION (provided by applicant): We propose a new approach to single-molecule DMA sequencing that can be done in a simple microtiter plate fitted with two electrodes. Over two-hundred different single DMA molecules would be sequenced simultaneously in a single well (maximum 10 nt/s per molecule) with read lengths 20 kb or more. Sequencing cost is potentially lower with this method compared to other long read approaches because of a high degree of multiplexing and because a simple microtiter plate substitutes for expensive micro or nanofabricated devices. While novel in concept, the proposed "electrokinetic" sequencing method would utilize modified polymerases and phosphate-labeled nucleotides from ongoing research on an alternative "charge-switch" method of single molecule sequencing. Ultrabright fluorescent nanoparticles and a prototype two-electrode sequencing chamber will be developed in Phase I. Particle transport kinetics will be characterized using the prototype chamber.