Patent ID: 11959137
Assignee: THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
Field: Organic fine chemistry (Chemistry)
Classification: CPC C | IPC C

Claim 19:
20. A method of sequencing, said method comprising:
contacting a primer oligonucleotide hybridized to a template polynucleotide with a first nucleotide analogue and a first polymerase thereby binding said first nucleotide analogue to a complementary nucleotide of template polynucleotide, wherein said first nucleotide analogue is associated with a detectable moiety and does not form a phosphodiester bond with the primer oligonucleotide;
detecting the detectable moiety;
removing the first nucleotide analogue; followed by
contacting the primer oligonucleotide with a second nucleotide analogue and a second polymerase thereby binding said second nucleotide analogue to a complementary nucleotide of said template polynucleotide and incorporating said second nucleotide analogue into the primer oligonucleotide, wherein the second nucleotide analogue is not associated with a detectable moiety, and wherein the first nucleotide analogue and the second nucleotide analogue comprise the same nucleobase type.