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US 11,142,789 B2Granted
Method of preparing libraries of template polynucleotides
AssigneeIllumina Cambridge Limited
InventorsNiall Anthony Gormley, Geoffrey Paul Smith, David Bentley, Roberto Rigatti, Shujun Luo
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Abstract
The present invention relates to a method for preparing a library of template polynucleotides and use thereof in methods of solid-phase nucleic acid amplification. More specifically, the invention relates to a method for preparing a library of template polynucleotides that have common sequences at their 5′ ends and at their 3′ ends.
Claims12
A method of generating a library of polynucleotide molecules, comprising:
- (a) providing a plurality of different polynucleotide duplexes;
- (b) providing identical forked polynucleotide adapters, wherein each adapter comprises a double-stranded annealed region and a mismatched single stranded region;
- (c) ligating the double-stranded annealed regions of the adapters to both ends of the polynucleotide duplexes to form adapter-target constructs;
- (d) annealing a single universal primer species to the mismatched regions of the adapter-target constructs;
- and
- (e) extending the primer to form extension products complementary to both strands of the adapter-target constructs,
- wherein the adapters are formed by annealing of partially complementary first and second polynucleotide strands, and at least one of the strands comprises a polynucleotide sequence complementary to the sequence of SEQ ID NO:
- 4.
5 more elements
Dependencies are inferred from the claim text.