Asymmetric adapter library construction
First Claim
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1. A method of producing an asymmetrically tagged nucleic acid fragment, said method comprising:
- i) ligating an adapter to each end of a double-stranded nucleic acid fragment, wherein said adapter comprises;
(a) a first and a second nucleic acid strand associated with each other via one or more complementary domains, said adapter having a first end and a second end;
(b) one or more region of substantial non-complementarity between said first and second nucleic acid strands;
(c) a ligation site positioned on said first end of said adapter configured to allow ligation of said adapter to said double stranded nucleic acid fragment; and
(d) a hairpin structure positioned on the 3′
strand on said second end of said adaptor, said hairpin structure producing a nucleic acid synthesis self-priming site; and
ii) performing a first round of nucleic acid synthesis initiated from said self-priming site using a nucleotide polymerase, thereby producing an asymmetrically tagged nucleic acid fragment.
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Abstract
The present invention provides methods and compositions for asymmetrically tagging a nucleic acid fragment using asymmetric adapters.
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Citations
17 Claims
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1. A method of producing an asymmetrically tagged nucleic acid fragment, said method comprising:
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i) ligating an adapter to each end of a double-stranded nucleic acid fragment, wherein said adapter comprises; (a) a first and a second nucleic acid strand associated with each other via one or more complementary domains, said adapter having a first end and a second end; (b) one or more region of substantial non-complementarity between said first and second nucleic acid strands; (c) a ligation site positioned on said first end of said adapter configured to allow ligation of said adapter to said double stranded nucleic acid fragment; and (d) a hairpin structure positioned on the 3′
strand on said second end of said adaptor, said hairpin structure producing a nucleic acid synthesis self-priming site; andii) performing a first round of nucleic acid synthesis initiated from said self-priming site using a nucleotide polymerase, thereby producing an asymmetrically tagged nucleic acid fragment. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 12, 13, 14, 16)
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9. A nucleic acid adapter comprising:
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(a) a first and a second nucleic acid strand associated with each other via one or more complementary domains, said adapter having a first end and a second end; (b) one or more region of substantial non-complementarity between said first and second nucleic acid strands; (c) a ligation site positioned on said first end of said adapter configured to allow ligation of said adapter to a double stranded nucleic acid fragment; and (d) a hairpin structure positioned on the 3′
strand on said second end of said adaptor, said hairpin structure producing a nucleic acid synthesis self-priming site. - View Dependent Claims (10, 11, 15, 17)
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Specification