Nanopores, methods for using same, methods for making same and methods for characterizing biomolecules using same

Abstract

Featured are devices and systems embodying one or more solid-state nanopores useable for sensing and / or characterizing single macromolecules as well as sequencing DNA or RNA and / or determining RNA secondary structures. In once solid state nanopore of the present invention the width and / or length of the nanopore is defined or established by sharp edges of cleaved crystals that are maintained in fixed relation during the formation of the insulating member including the nanopore. In another aspect of the present invention, there is featured a linear or 2-D electrically addressable array of nanopores, where the nanopores are located at points of intersections between grooves formed in an upper surface of the insulating member and a groove formed in a lower surface of the insulating member.

Description

FIELD OF INVENTION [0001] The present invention relates to devices used for characterizing biomolecules as well as sequencing DNA and RNA, and more particularly to a nanopore and / or an array of nanopores for use in rapid characterization of biomolecules and high through-put DNA sequencing. BACKGROUND OF THE INVENTION [0002] Transport of nucleic acids and other biological molecules through membrane channels of nanometer scale is ubiquitous in Nature. Examples include the movement of RNA molecules and transcription factors across nuclear pores; viral DNA injected through a bacterium membrane in phage infection; and the uptake of specific oligonucleotides by membrane proteins. Understanding the function of these nanoscale channels and how biomolecules move through them is a major task of modem molecular biology and biophysics. In recent years, a fascinating field of nanopore biophysics has emerged in the interdisciplinary area of molecular biology, nanotechnology, and single molecule ...

AI Technical Summary

Problems solved by technology

However, because the natural nanopores are long channels, typically 30 nm in length, many nucleotides are inside the channel at any time during translocation and the effect of individual nucleotides on the ionic conductance is lost.

In fact, the physics of how a linear flexible molecule (such as a DNA) threads through a nanopore under the influence of a driving force is poorly understood, making it difficult to extract size information from the ionic current data.

Previous temperature-dependent measurements of DNA translocation through α-hemolysin pore suggest that thermal motion of DNA is a source of noise in the blockage current.

All these processes except optical trapping are stochastic and can be very time consuming.

In general these techniques have been used to validate that a nanopore can be used in the described fashion, but have not been used to yield a solid state nanopore device having high through-put.

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