HYBRIDIZATION ASSISTED NANOPORE SEQUENCING
First Claim
1. A method for determining the sequence of a biomolecule strand of interest, comprising the steps of:
- providing a sequencing apparatus having a first fluid chamber, a second fluid chamber, a membrane positioned between said first and second chambers and a nanopore extending through said membrane such that said first and second chambers are in fluid communication via said nanopore;
providing a single-stranded biomolecule;
providing a first plurality of matching probes having a known sequence;
hybridizing said first plurality of probes with said single-stranded biomolecule such that said first plurality of probes attach to portions of said single-stranded biomolecule to produce a partially hybridized biomolecule;
introducing said partially hybridized biomolecule into said first chamber;
translocating said partially hybridized biomolecule from said first chamber through said nanopore and into said second chamber;
monitoring changes in current across said nanopore as said partially hybridized biomolecule is translocated therethrough, said changes in electrical potential corresponding to locations along said partially hybridized biomolecule containing one of said first plurality of probes; and
recording said changes in electrical potential as a function of time.
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Accused Products
Abstract
A method of employing a nanopore structure in a manner that allows the detection of the positions (relative and/or absolute) of nucleic acid probes that are hybridized onto a single-stranded nucleic acid molecule. In accordance with the method the strand of interest is hybridized with a probe having a known sequence. The strand and hybridized probes are translocated through a nanopore. The fluctuations in current measured across the nanopore will vary as a function of time corresponding to the passing of a probe attachment point along the strand. These fluctuations in current are then used to determine the attachment positions of the probes along the strand of interest. This probe position data is then fed into a computer algorithm that returns the sequence of the strand of interest.
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Citations
19 Claims
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1. A method for determining the sequence of a biomolecule strand of interest, comprising the steps of:
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providing a sequencing apparatus having a first fluid chamber, a second fluid chamber, a membrane positioned between said first and second chambers and a nanopore extending through said membrane such that said first and second chambers are in fluid communication via said nanopore;
providing a single-stranded biomolecule;
providing a first plurality of matching probes having a known sequence;
hybridizing said first plurality of probes with said single-stranded biomolecule such that said first plurality of probes attach to portions of said single-stranded biomolecule to produce a partially hybridized biomolecule;
introducing said partially hybridized biomolecule into said first chamber;
translocating said partially hybridized biomolecule from said first chamber through said nanopore and into said second chamber;
monitoring changes in current across said nanopore as said partially hybridized biomolecule is translocated therethrough, said changes in electrical potential corresponding to locations along said partially hybridized biomolecule containing one of said first plurality of probes; and
recording said changes in electrical potential as a function of time. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification