HYBRID NANOPORE DEVICE WITH OPTICAL DETECTION AND METHODS OF USING SAME

摘要

The invention is directed to a device comprising a protein nanopore immobilized in a lipid layer within an aperture of a solid phase substrate, which provides a stable platform for using first and second members of one or more FRET pairs to generate optical signals as a labeled analyte translocates through the bore of the protein nanopore. In another aspect, the invention is directed to the use of the device to determine the nucleotide sequence of a polynucleotide analyte.

主权项

1. A device for detecting an analyte, the device comprising:
a solid phase membrane separating a first chamber and a second chamber, the solid phase membrane having at least one aperture with a wall connecting the first chamber and the second chamber a lipid layer disposed on the at least one surface of the solid phase membrane; a protein nanopore immobilized in the aperture, the protein nanopore having a bore and interacting with the lipid layer to form a seal with the solid phase membrane in the aperture so that fluid, communication between the first chamber and the second chamber occurs solely through the bore of the protein nanopore; and wherein the solid phase membrane or the protein nanopore has attached thereto at least one first member of a fluorescent resonance energy transfer (FRET) pair, so that whenever an analyte having at least one second member of the FRET pair attached thereto traverses the bore, the second member passes with a FRET distance of the first member of the FRET pair.