Polynucleotide modification methods
First Claim
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1. A method for modifying a template double stranded polynucleotide, comprising:
- (a) contacting a double stranded template polynucleotide with a MuA transposase and a population of double stranded MuA substrates each comprising at least one overhang of universal nucleotides, such that the transposase fragments the template polynucleotide into two or more double-stranded fragments, and ligates a substrate to an end of each of the double stranded fragments, thereby producing a plurality of fragment/substrate constructs; and
(b) ligating the overhangs to the fragments in the constructs and thereby producing a plurality of modified double stranded polynucleotides.
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Abstract
The invention relates to a method for modifying a template double stranded polynucleotide, especially for characterization using nanopore sequencing. The method produces from the template a plurality of modified double stranded polynucleotides. These modified polynucleotides can then be characterized.
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Citations
14 Claims
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1. A method for modifying a template double stranded polynucleotide, comprising:
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(a) contacting a double stranded template polynucleotide with a MuA transposase and a population of double stranded MuA substrates each comprising at least one overhang of universal nucleotides, such that the transposase fragments the template polynucleotide into two or more double-stranded fragments, and ligates a substrate to an end of each of the double stranded fragments, thereby producing a plurality of fragment/substrate constructs; and (b) ligating the overhangs to the fragments in the constructs and thereby producing a plurality of modified double stranded polynucleotides. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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Specification