THIOLATED NUCLEOTIDE ANALOGUES FOR NUCLEIC ACID SYNTHESIS

摘要

The present disclosure provide systems, compositions, methods, reagents, kits and products for extending a nucleic acid that includes incorporating a nucleotide residue at a terminus of a nucleic acid using a polymerase enzyme and at least one nucleotide, wherein the at least one nucleotide includes a thiophosphate moiety, and wherein the at least one nucleotide is resistant to hydrolysis by phosphatase. In some embodiments, the nucleotide incorporation can be conducted in the presence of a phosphatase. In some embodiments, the nucleotide incorporation can be conducted in the presence of at least on chelation moiety that is configured to bind an orthophosphate moiety.

主权项

1. A method of identifying a base at a position in a target nucleic acid, comprising:
a) incorporating a nucleotide at a terminus of an extension primer that is hybridized to the target nucleic acid using a polymerase enzyme and at least one nucleotide, wherein the terminal phosphate of the at least one nucleotide includes a thio-phosphate moiety, wherein the at least one nucleotide is resistant to hydrolysis by a phosphatase enzyme, and wherein the nucleotide incorporation produces a thio-pyrophosphate and a hydrogen ion or a proton; and b) identifying the nucleotide that is incorporated at the terminus of the extension primer.