Patent ID: 8163487

Claim:
A method of extending a primer nucleic acid, the method comprising: incubating a template nucleic acid with at least one nucleotide incorporating biocatalyst, at least one 2′-terminator nucleotide, and at least one primer nucleic acid that is at least partially complementary to at least a subsequence of the template nucleic acid, which primer nucleic acid comprises DNA, under conditions whereby the nucleotide incorporating biocatalyst extends the primer nucleic acid to produce at least one extended primer nucleic acid by incorporating the 2′-terminator nucleotide at a terminal end of the extended primer nucleic acid, wherein the 2′-terminator nucleotide comprises the formula: wherein R 1 is H, OH, a hydrophilic group, or a hydrophobic group; B is at least one homocyclic ring, at least one heterocyclic ring, at least one aryl group, or combinations thereof; BG is a blocking group; and Z is O or CH 2 ; and represents a single or double bond, wherein BG comprises the formula: wherein X is O, S, NR 3 , CR 3 R 4 or SiR 3 R 4 ; Y is CR 5 R 6 R 7 , SiR 5 R 6 R 7 , OR 5 , SR 5 , or NHR 5 ; R 2 is H, OH, NHR 8 , SR 8 , an alkyl group, a benzyl group, an aryl group, an alkenyl group, an alkynyl group, an alkoxy group, or combinations thereof; and R 3 , R 4 , R 5 , R 6 , R 7 , and R 8 are independently selected from H, an alkyl group, a benzyl group, an aryl group, an alkenyl group, an alkynyl group, or combinations thereof; or wherein X is CR 3 R 4 R 5 , SiR 3 R 4 R 5 , OR 3 , SR 3 , or NHR 3 ; R 2 is H, OH, NHR 6 , SR 6 , an alkyl group, a benzyl group, an aryl group, an alkenyl group, an alkynyl group, an alkoxy group, or combinations thereof; and, R 3 ,R 4 , R 5 , and R 6 are independently selected from H, an alkyl group, a benzyl group, an aryl group, an alkenyl group, an alkynyl group, or combinations thereof.