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US 11,236,385 B2Granted
Method for characterising a double stranded nucleic acid using a nano-pore and anchor molecules at both ends of said nucleic acid
AssigneeOxford Nanopore Technologies PLC
InventorsMarion Louise Crawford, James White
Life of this patent
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Abstract
Method for characterising a double stranded nucleic acid using a nano-pore and anchor molecules at both ends of said nucleic acid.
Claims18
A method for modifying a target polynucleotide for characterisation using a transmembrane pore in a membrane, comprising:
- (a) ligating a Y adaptor to one end of the target polynucleotide;
- and
- (b) attaching to the Y adaptor one or more first anchors for coupling the target polynucleotide to the membrane and attaching directly to the target polynucleotide, one or more second anchors for coupling the polynucleotide to the membrane and thereby providing a modified target polynucleotide;
- wherein the strength of coupling of the one or more second anchors to the membrane is greater than the strength of coupling of the one or more first anchor to the membrane.
1 more element
A method of characterising a target polynucleotide using a transmembrane pore in a membrane, comprising:
- a) providing the target polynucleotide with a Y adaptor at one end, wherein the Y adaptor comprises one or more first anchors for coupling the polynucleotide to the membrane, wherein the target polynucleotide comprises one or more second anchors for coupling the polynucleotide to the membrane and wherein the strength of coupling of the one or more second anchors is greater than the strength of coupling of the one or more first anchors to the membrane;
- b) contacting the polynucleotide provided in step a) with the transmembrane pore such that at least one strand of the polynucleotide moves through the pore;
- and
- c) taking one or more measurements as the at least one strand of the polynucleotide moves with respect to the pore wherein the measurements are indicative of one or more characteristics of the at least one strand of the polynucleotide and thereby characterising the target polynucleotide.
1 more element
A method of characterising a target polynucleotide using a transmembrane pore in a membrane, comprising:
- a) providing the target polynucleotide with a Y adaptor at one end, wherein the Y adaptor comprises one or more first anchors for coupling the polynucleotide to the membrane, wherein the polynucleotide comprises one or more second anchors for coupling the polynucleotide to the membrane and wherein the strength of coupling of the one or more second anchors to the membrane is greater than the strength of coupling of the one or more first anchors to the membrane;
- b) contacting the polynucleotide provided in step a) with the transmembrane pore, a polymerase and labelled nucleotides such that phosphate labelled species are sequentially released when nucleotides are added to the polynucleotide by the polymerase, wherein the phosphate species contain a label specific for each nucleotide;
- and
- c) detecting the phosphate labelled species using the pore and thereby characterising the target polynucleotide.
1 more element
Dependencies are inferred from the claim text.