	US 12,110,515 B2
	*Modified Thermococcus* polymerases**
Souad Naji, La Jolla, CA (US); Carl W. Fuller, Berkley Heights, NJ (US); Eli N. Glezer, Del Mar, CA (US); and Andrew Spaventa, San Diego, CA (US)
Assigned to Singular Genomics Systems, Inc., San Diego, CA (US)
Filed by Singular Genomics Systems, Inc., San Diego, CA (US)
Filed on Oct. 20, 2022, as Appl. No. 18/048,388.
Application 18/048,388 is a continuation of application No. 17/017,578, filed on Sep. 10, 2020, granted, now 11,512,295.
Claims priority of provisional application 62/899,644, filed on Sep. 12, 2019.
Prior Publication US 2023/0227798 A1, Jul. 20, 2023
Int. Cl. C12N 9/12 (2006.01); C12N 15/10 (2006.01); C12Q 1/6874 (2018.01)

CPC C12N 9/1252 (2013.01) [C12N 15/102 (2013.01); C12Q 1/6874 (2013.01); C12Y 207/07007 (2013.01)]

18 Claims

1. A method of extending a primer, said method comprising contacting a primer hybridized to a template polynucleotide with a nucleotide and extending said primer with a polymerase by incorporating the modified nucleotide to form an extended primer, wherein the polymerase comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 1; and further comprises:

an alanine at amino acid position 411 or an amino acid functionally equivalent to amino acid position 411;

a glycine at amino acid position 412 or an amino acid functionally equivalent to amino acid position 412; and

a proline at amino acid position 413 or an amino acid functionally equivalent to amino acid position 413; and

an alanine or glycine at amino acid position 93 or an amino acid functionally equivalent to amino acid position 93.