US 11,788,138 B2
Nucleic acid sequencing using affinity reagents
Radoje Drmanac, Los Altos Hills, CA (US); Snezana Drmanac, Los Altos Hills, CA (US); Handong Li, San Jose, CA (US); Xun Xu, Shenzhen (CN); Matthew J. Callow, Redwood City, CA (US); Leon Eckhardt, Soquel, CA (US); and Naibo Yang, Burlingame, CA (US)
Assigned to MGI Tech Co., Ltd., Shenzhen (CN); and BGI Shenzhen, Shenzhen (CN)
Filed by MGI Tech Co., Ltd., Shenzhen (CN); and BGI Shenzhen, Shenzhen (CN)
Filed on Oct. 29, 2020, as Appl. No. 17/84,482.
Application 17/084,482 is a continuation of application No. 15/862,566, filed on Jan. 4, 2018, granted, now 10,851,410.
Claims priority of provisional application 62/490,511, filed on Apr. 26, 2017.
Claims priority of provisional application 62/442,263, filed on Jan. 4, 2017.
Prior Publication US 2021/0254150 A1, Aug. 19, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6874 (2018.01); C12Q 1/6876 (2018.01); C07K 16/44 (2006.01); C12Q 1/6804 (2018.01); A61K 47/00 (2006.01); C12Q 1/6869 (2018.01); C07K 16/28 (2006.01)
CPC C12Q 1/6874 (2013.01) [A61K 47/00 (2013.01); C07K 16/2851 (2013.01); C07K 16/44 (2013.01); C12Q 1/6804 (2013.01); C12Q 1/6869 (2013.01); C12Q 1/6876 (2013.01); C07K 2317/73 (2013.01)] 47 Claims
 
1. A method for identifying a nucleobase of a non-labeled reversible terminator (NLRT) deoxyribonucleotide incorporated at a 3′ terminus of a primer extension product, wherein the NLRT deoxyribonucleotide comprises a nucleobase, a sugar moiety, and a cleavable blocking group, said method comprising
(a) combining (i) the primer extension product including the incorporated NLRT deoxyribonucleotide and (ii) a first affinity reagent that binds to the incorporated NLRT deoxyribonucleotide, wherein the first affinity reagent binds to the nucleobase, the cleavable blocking group, or both, and
(b) detecting binding of the first affinity reagent to the incorporated NLRT deoxyribonucleotide, wherein binding of the first affinity reagent identifies a nucleobase associated with the incorporated NLRT deoxyribonucleotide.