NANOSENSORS USING FLIGHT-TIME IDENTIFICATION OF MONONUCLEOTIDES FOR SINGLE-MOLECULE DNA SEQUENCING
Steven A. Soper1, Sunggook Park2, Elizabeth Podlaha-Murphy3
1University of North Carolina Chapel Hill, USA;
2Louisiana State University, USA;
3Northeastern University, USA
We are generating a single-molecule DNA sequencing platform that can acquire sequencing information with high accuracy that is built from a single-molecule sequencing process. The technology employs high density arrays of nanosensors that read the identity of individual mononucleotides from their characteristic flight-time through a 2-dimensional (2D) nanochannel (~20 nm in width and depth; >100 μm in length) fabricated in a plastic via nano-imprinting (NIL). The mononucleotides are generated from an intact DNA fragment using a highly processive exonuclease, which is covalently anchored to a solid support contained within a bioreactor that sequentially feeds mononucleotides into the 2D nanochannel. The identity of the mononucleotides is deduced from a moleculardependent flight-time through the 2D nanochannel. The flight time is read in a label-less fashion by measuring current transients induced by a single mononucleotide when it travels through a constriction with molecular dimensions (<10 nm in diameter) poised at the input/ output ends of the flight tube. In this presentation, our efforts on building these nanosensors using NIL will be discussed and the detection of single molecules using electrical transduction with their identity deduced from the associated flight time. Finally, information on the manipulation of single DNA molecules using nanofluidic circuits will be discussed that takes advantage of forming unique nano-scale features to shape electric fields for DNA manipulation.