Method
AssigneeOxford Nanopore Technologies PLC
InventorsJames Edward Graham, Etienne Raimondeau, Rebecca Victoria Bowen
Life of this patent
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Abstract
A method for selectively modifying a target polynucleotide in a sample of polynucleotides, the method comprising contacting a sample of polynucleotides with a guide polynucleotide that binds to a sequence in the target polynucleotide and a polynucleotide-guided effector protein such that the polynucleotide-guided effector protein cuts the target polynucleotide to produce a cut end comprising an overhang; and attaching an adapter to the cut end in the target polynucleotide.
Claims30
A method for selectively adapting a target polynucleotide in a sample of polynucleotides, the method comprising:
- (a) protecting the ends of the polynucleotides in the sample;
- (b) contacting the polynucleotides with a guide polynucleotide that binds to a sequence in the target polynucleotide and a polynucleotide-guided effector protein such that the polynucleotide-guided effector protein cuts the target polynucleotide to produce two opposing cut ends at a site determined by the sequence to which the guide polynucleotide binds;
- and
- attaching an adapter to one or both of the two opposing cut ends in the target polynucleotide, wherein the adapter attaches to one or both of the cut ends in the target polynucleotide but does not attach to the protected ends of the polynucleotides in the sample.
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A kit for selectively modifying a target polynucleotide in a sample of polynucleotides, the kit comprising a dephosphorylase, an adapter comprising a single N or polyN tail, wherein N is the nucleotide A, T, C or G, and optionally one or more of a polymerase, a ligase, a polynucleotide-guided effector protein and a guide polynucleotide.
A method for selectively adapting a target polynucleotide in a sample of polynucleotides, the method comprising:
- (a) contacting the polynucleotides in the sample with two guide polynucleotides that bind to a sequences in the target polynucleotide and a polynucleotide-guided effector protein, wherein the sequences to which the two guide polynucleotides bind direct the polynucleotide-guided effector protein to two sites, such that the polynucleotide-guided effector protein cuts the target polynucleotide at least one of the two sites to produce two opposing cut ends;
- and
- (b) attaching an adapter to one or both of the two opposing cut ends in the target polynucleotide.
Dependencies are inferred from the claim text.