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 12:
13. A method of sequencing a template polynucleotide, said method comprising:
(i) contacting a primer hybridized to the template polynucleotide with a first nucleotide analogue comprising a first reversible terminator and extending said primer with a first polymerase by incorporating the first nucleotide analogue to form an extended primer;
(ii) contacting the extended primer with a second nucleotide analogue capable of forming at least one hydrogen bond with a complementary nucleotide of the template polynucleotide, wherein the second nucleotide analogue is associated with a detectable moiety; detecting the detectable moiety; wherein the second nucleotide analogue is not incorporated into the extended primer;
(iii) and contacting the extended primer with a third nucleotide analogue comprising a second reversible terminator and extending said extended primer with a polymerase by incorporating the third nucleotide analogue, wherein said first reversible terminator and said second nucleotide analogue are removed prior to step (iii); and
(iv) repeating steps (i), (ii), and (iii), thereby sequencing the template polynucleotide.