title: DNA sequencing by nanopore-induced photon emission.
abstract: Nanopore-based DNA analysis is an extremely attractive area of research due to the simplicity of the method, and the ability to not only probe individual molecules, but also to detect very small amounts of genomic material. Here, we describe the materials and methods of a novel, nanopore-based, single-molecule DNA sequencing system that utilizes optical detection. We convert target DNA according to a binary code, which is recognized by molecular beacons with two types of fluorophores. Solid-state nanopores are then used to sequentially strip off the beacons, leading to a series of photon bursts that can be detected with a custom-made microscope. We do not use any enzymes in the readout stage; thus, our system is not limited by the highly variable processivity, lifetime, and inaccuracy of individual enzymes that can hinder throughput and reliability. Furthermore, because our system uses purely optical readout, we can take advantage of high-end, wide-field imaging devices to record from multiple nanopores simultaneously. This allows an extremely straightforward parallelization of our system to nanopore arrays.
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA, USA.
authors.forename: Alon
authors.initials: A
authors.lastname: Singer
authors.forename: Ben
authors.initials: B
authors.lastname: McNally
authors.forename: Ruby Dela
authors.initials: RD
authors.lastname: Torre
authors.forename: Amit
authors.initials: A
authors.lastname: Meller
mesh_terms: D015342:DNA Probes / Q000737:chemistry; D004270:DNA, Circular / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D046228:Microarray Analysis / Q000379:methods; D046529:Microscopy, Electron, Transmission; D058608:Nanopores; D017785:Photons; D017422:Sequence Analysis, DNA / Q000379:methods*; D017655:Silicon Compounds / Q000737:chemistry
chemical_list: D015342:DNA Probes; D004270:DNA, Circular; D005456:Fluorescent Dyes; D017655:Silicon Compounds; C032734:silicon nitride