Template switch-based methods for producing a product nucleic acid
AssigneeClontech Laboratories, Inc.
InventorsCraig Betts, Andrew Alan Farmer
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Abstract
Provided are methods of producing a product nucleic acid. The methods include combining a template deoxyribonucleic acid (DNA), a polymerase, a template switch oligonucleotide, and dNTPs into a reaction mixture. The components are combined into the reaction mixture under conditions sufficient to produce a product nucleic acid that includes the template DNA and the template switch oligonucleotide each hybridized to adjacent regions of a single product nucleic acid that includes a region polymerized from the dNTPs by the polymerase. Aspects of the invention further include compositions and kits.
Claims15
A method comprising:
- combining:
- a template deoxyribonucleic acid (DNA);
- a retroviral reverse transcriptase;
- a template switch oligonucleotide;
- a first DNA primer comprising a 3′ domain that hybridizes to the template DNA;
- and
- dNTPs;
- into a reaction mixture under conditions sufficient to produce a product nucleic acid, wherein the product nucleic acid comprises the template DNA and the template switch oligonucleotide each hybridized to adjacent regions of a single hybrid nucleic acid comprising a region polymerized from the dNTPs by the retroviral reverse transcriptase, wherein the region polymerized from the dNTPs comprises a domain that is complementary to and hybridized to the 5′ end of the template DNA.
Dependencies are inferred from the claim text.