US 11,851,687 B2
Modified Pyrococcus polymerases and uses thereof
Souad Naji, La Jolla, CA (US); Eli N. Glezer, Del Mar, CA (US); Youngjin Cho, San Diego, CA (US); Zachary Terranova, San Diego, CA (US); and Abrehet Abdu, 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 Mar. 19, 2021, as Appl. No. 17/207,387.
Application 17/207,387 is a division of application No. 16/803,763, filed on Feb. 27, 2020, granted, now 11,034,942.
Prior Publication US 2021/0284975 A1, Sep. 16, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/12 (2006.01); C12N 15/63 (2006.01); C12Q 1/68 (2018.01); C12Q 1/6874 (2018.01)
CPC C12N 9/1252 (2013.01) [C12N 15/63 (2013.01); C12Q 1/6874 (2013.01); C12Y 207/07007 (2013.01)] 17 Claims
 
1. A method of sequencing a nucleic acid sequence comprising:
a) hybridizing a nucleic acid template with a primer to form a primer-template hybridization complex;
b) contacting the primer-template hybridization complex with a DNA polymerase and nucleotides, wherein each of the nucleotides comprises a detectable label;
c) subjecting the primer-template hybridization complex to conditions which enable the polymerase to incorporate a nucleotide into the primer-template hybridization complex to form a modified primer-template hybridization complex;
d) detecting the detectable label; thereby sequencing a nucleic acid sequence;
wherein the DNA polymerase comprises an amino acid sequence that is at least 80% identical to a continuous 500 amino acid sequence within SEQ ID NO: 1; comprising the following amino acids:
an alanine at amino acid position 409 or at an amino acid position functionally equivalent to amino acid position 409;
a glycine at amino acid position 410 or at an amino acid position functionally equivalent to amino acid position 410;
a proline at amino acid position 411 or at an amino acid position functionally equivalent to amino acid position 411.