Preparation of templates for nucleic acid sequencing
First Claim
1. A method comprising,(i) amplifying a template using amplification primers on a solid support to produce a plurality of double-stranded nucleic acid molecules such that both strands of each double-stranded nucleic acid molecule are attached to the solid support at their 5′
- ends, wherein a subset of the amplification primers comprise a cleavage site and wherein the cleavage site is positioned in a double-stranded region of each double-stranded molecule; and
(ii) cleaving only one strand of the double stranded molecules at the cleavage site to produce a cleaved strand and an un-cleaved strand.
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Abstract
The invention relates to methods of generating templates for a nucleic acid sequencing reaction which comprise: providing at least one double-stranded nucleic acid molecule, wherein both strands of the double-stranded nucleic acid molecule are attached to a solid support at the 5′ end, cleaving one or both strands of the double-stranded nucleic acid molecule, and subjecting the cleaved strand(s) to denaturing conditions to remove the portion of the cleaved strand(s) not attached to the solid support, thereby generating a partially or substantially single-stranded template for a nucleic acid sequencing reaction.
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Citations
20 Claims
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1. A method comprising,
(i) amplifying a template using amplification primers on a solid support to produce a plurality of double-stranded nucleic acid molecules such that both strands of each double-stranded nucleic acid molecule are attached to the solid support at their 5′ - ends, wherein a subset of the amplification primers comprise a cleavage site and wherein the cleavage site is positioned in a double-stranded region of each double-stranded molecule; and
(ii) cleaving only one strand of the double stranded molecules at the cleavage site to produce a cleaved strand and an un-cleaved strand. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
- ends, wherein a subset of the amplification primers comprise a cleavage site and wherein the cleavage site is positioned in a double-stranded region of each double-stranded molecule; and
Specification