Continuous extension and deblocking in reactions for nucleic acids synthesis and sequencing
US10378051B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 26, 2012 |
| Grant date | Aug 13, 2019 |
| Priority date | — |
| Expiry date | Jan 28, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6874
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A reaction mixture including (a) a nucleic acid having a primer hybridized to a template, (b) nucleotide analogs, wherein the nucleotide analogs have a blocking moiety; (c) a polymerase that is capable of forming an extended primer by adding the nucleotide analogs to the primer, and (d) a deblocking agent that is capable of removing the blocking moiety from the extended primer. Also provided is a method of synthesizing a polynucleotide including sequentially adding a plurality of the different nucleotides analogs to the nucleic acid via several reaction cycles in the reaction mixture, wherein each reaction cycle includes (i) the polymerase adding a nucleotide analog to the nucleic acid to form a transient nucleic acid species comprising a blocking moiety, and (ii) the deblocking agent modifying the transient nucleic acid species to remove the blocking moiety.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.