Back to results
US 2019/0055592 A1Published application
Nanopore Discrimination of Target Polynucleotides from Sample Background by Fragmentation and Payload Binding
AssigneeTwo Pore Guys, Inc.
InventorsTrevor J. Morin, William B. Dunbar, Daniel A. Heller, Tyler Shropshire
Life of this patent
Estimating the term…
Abstract
Disclosed herein are methods and compositions for detecting a target DNA sequence from a sample that does not require sample purification or amplification. The method uses fragmentation, sequence-specific binding or ligation of probes, and payload molecules for selective detection of the target-sequence using a nanopore sensor.
Claims73
A method of detecting the presence or absence of a target polynucleotide sequence suspected to be present in a sample, comprising:
- a) fragmenting polynucleotides in a sample suspected of comprising a target polynucleotide comprising a target sequence;
- b) contacting said sample with a probe adapted to bind specifically to a fragmented target polynucleotide comprising said target sequence under conditions that promote binding of said probe to said fragmented polynucleotide to form a polynucleotide-probe complex;
- c) loading said sample into a nanopore device comprising a nanopore, a first chamber, and a second chamber, wherein said first and second chamber are in electrical and fluidic communication through said nanopore via a conducting fluid, and wherein said nanopore device further comprises a sensor configured to identify objects passing through the nanopore;
- d) applying an electrical potential across said nanopore to induce translocation of said polynucleotide or polynucleotide-probe complex through said nanopore;
- and
- e) detecting an electrical signal associated with the translocation of said polynucleotide or polynucleotide-probe complex through the nanopore.
3 more elements
A method of quantifying a target polynucleotide sequence in a sample, comprising:
- a) fragmenting polynucleotides in a sample suspected of comprising a target polynucleotide comprising a target sequence;
- b) contacting said sample with a probe adapted to bind specifically to a fragmented target polynucleotide comprising said target sequence under conditions that promote binding of said probe to said fragmented polynucleotide to form a polynucleotide-probe complex;
- c) loading said sample into a nanopore device comprising a nanopore, a first chamber, and a second chamber, wherein said first and second chamber are in electrical and fluidic communication through said nanopore via a conducting fluid, and wherein said nanopore device further comprises a sensor configured to identify objects passing through the nanopore;
- d) applying an electrical potential across said nanopore to induce translocation of said polynucleotide or polynucleotide-probe complex through said nanopore;
- e) detecting an electrical signal associated with the translocation of said polynucleotide or polynucleotide-probe complex through the nanopore;
- and
- f) analyzing said electrical signal to quantify said target polynucleotide sequence in said sample.
4 more elements
A kit comprising:
- a device comprising a nanopore, wherein said nanopore separates an interior space of the device into two volumes, wherein the device comprises a sensor for said nanopore adapted to identify objects passing through the nanopore;
- a probe adapted to bind specifically to a fragmented target polynucleotide comprising a target sequence;
- and
- instructions for use to detect the presence or absence of said target sequence in a sample.
1 more element
Dependencies are inferred from the claim text.