Methods for Adding Adapters to Nucleic Acids and Compositions for Practicing the Same
AssigneeTakara Bio USA, Inc.
InventorsCraig Betts, Andrew Alan Farmer
Life of this patent
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Abstract
Provided are methods of adding adapters to nucleic acids. The methods include combining in a reaction mixture a template nucleic acid, a template switch oligonucleotide, a polymerase, and dNTPs. The reaction mixture components are combined under conditions sufficient to produce a product nucleic acid that includes the template nucleic acid and the template switch oligonucleotide each hybridized to adjacent regions of a single product nucleic acid including a region polymerized from the dNTPs by the polymerase. The methods further include attaching sequencing platform adapter constructs to ends of the product nucleic acid or a derivative thereof. Aspects of the invention further include compositions and kits.
Claims37
1-50. (canceled)
A kit comprising:
- a template switch oligonucleotide comprising a 3’ hybridization domain and one or more modified nucleotides 5′ of the 3′ hybridization domain;
- and
- a sequencing platform adapter construct.
A kit comprising:
- a template switch oligonucleotide comprising one or more modified nucleotides comprising a modification selected from the group consisting of:
- a 5′ amino end modification, a 3′ amino end modification, a biotin end modification, a digoxigenin (DIG) end modification, a 5′ phosphate end modification, a 3′ phosphate end modification a thiol end modification, a dye end modification, a quencher end modification, a locked nucleic acid (LNA), a 2′-fluoro-arabino nucleic acid (FANA) modification, a 2′-O-methyl RNA modification, a 2′-fluoro RNA modification, a phosphorothioate linkage modification, a 3′-3′ reversed linkage modification, and a 5′-5′ reversed linkage modification;
- and
- a sequencing platform adapter construct.
A kit comprising:
- a template switch oligonucleotide comprising a sequencing platform adapter construct;
- and
- a template switching polymerase.
Dependencies are inferred from the claim text.