Method for selecting polynucleotides based on enzyme interaction duration
AssigneeOxford Nanopore Technologies PLC
InventorsAndrew John Heron, Rebecca Victoria Bowen
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Abstract
A method for selecting polynucleotides, the method comprising: allowing a nucleic acid handling enzyme to move along multiple polynucleotides in a sample for a defined time period, wherein the enzyme is loaded onto each of the multiple polynucleotides and wherein one or more molecule of the enzyme moves along each of the multiple polynucleotides; and selecting polynucleotides based on whether or not the enzyme reaches the end of and/or unbinds from the polynucleotides in the defined time period.
Claims24
A method for selecting polynucleotides for characterizing that are at least 1 kb in length, the method comprising:
- (i) allowing a plurality of nucleic acid handling enzymes to move along multiple polynucleotides in a sample for a defined time period, wherein the enzymes of the plurality are loaded onto each of the multiple polynucleotides and wherein one or more of the enzymes moves along each of the multiple polynucleotides;
- (ii) selecting polynucleotides from step (i) that are at least 1 kb in length based on whether or not the enzymes reach the end of and/or unbind from the polynucleotides in the defined time period;
- (iii) physically separating the selected polynucleotides from unselected polynucleotides having a length less than 1 kb prior to contacting the selected polynucleotides with a transmembrane pore in step (iv), wherein the physical separation does not comprise gel electrophoresis;
- and
- (iv) characterizing the selected polynucleotides, wherein the characterizing step (iv) comprises (a) contacting the selected polynucleotides with the transmembrane pore.
Dependencies are inferred from the claim text.