US 12,031,179 B2
Methods and compositions for reducing nucleotide impurities
Abrehet Abdu, San Diego, CA (US); Eli N. Glezer, Del Mar, CA (US); Laura Ann Hale, Del Mar, CA (US); and Jiawen Li, San Diego, CA (US)
Assigned to Singular Genomics Systems, Inc., San Diego, CA (US)
Filed by SINGULAR GENOMICS SYSTEMS, INC., La Jolla, CA (US)
Filed on Nov. 1, 2021, as Appl. No. 17/516,554.
Claims priority of provisional application 63/108,179, filed on Oct. 30, 2020.
Prior Publication US 2022/0136048 A1, May 5, 2022
Int. Cl. C12Q 1/6869 (2018.01); B01L 3/00 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6874 (2018.01)
CPC C12Q 1/6874 (2013.01) [B01L 3/502715 (2013.01); C12Q 1/6806 (2013.01); C12Q 2600/166 (2013.01)] 17 Claims
OG exemplary drawing
 
1. A composition comprising:
(a) labeled nucleotide triphosphates comprising a free 3′-OH,
(b) labeled nucleotide triphosphates comprising a 3′ reversible terminator moiety, and
(c) a non-immobilized depletion polynucleotide comprising a depletion primer hybridized to a depletion template; and
(d) a depletion polymerase that is active to selectively incorporate the nucleotide triphosphates comprising a free 3′-OH into the depletion primer and incapable of incorporating the labeled nucleotide triphosphates comprising a 3′ reversible terminator moiety;
(e) a sequencing primer hybridized to an immobilized polynucleotide; and
(f) a sequencing polymerase capable of incorporating the nucleotide triphosphates comprising a free 3′-OH and capable of incorporating the labeled nucleotide triphosphates comprising a 3′ reversible terminator moiety.