Granted Patent B2

US 12,215,124 · App. 16/969,879 · Granted Feb 4, 2025

Inventors: Ronald Graham (Carlsbad, CA); Andrew Spaventa (La Jolla, CA); Eli Glezer (Del Mar, CA)

Assignee: Singular Genomics Systems, Inc.

C07H21/04C12Q1/6869C12Q2600/16

Abstract

Claims (28)

1. A nucleotide analogue having the formula:

wherein

B is a base or analogue thereof;

L 2B , L 2C , L 2D , and L 2E are independently a bond, —NN—, —NHC(O)—, —C(O)NH—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene, wherein at least one of L 2A , L 2B , L 2C , L 2D , and L 2E is not a bond;

L 4 is a bond, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene;

R 4 is substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R 5 is a detectable label, an anchor moiety, or an affinity anchor moiety; and

X 1 is hydrogen, halogen, —N 3 , or —CN.

2. The nucleotide analogue of claim 1 , having the formula:

3. The nucleotide analogue of claim 1 , wherein X 1 is —F.

4. The nucleotide analogue of claim 1 , wherein X 1 is hydrogen.

5. The nucleotide analogue of claim 1 , wherein X 1 is —Cl.

6. The nucleotide analogue of claim 1 , wherein X 1 is —Br.

7. The nucleotide analogue of claim 1 , wherein X 1 is —I.

8. The nucleotide analogue of claim 1 , wherein L 4 is substituted or unsubstituted C 1 -C 8 alkylene, substituted or unsubstituted 2 to 8 membered heteroalkylene, substituted or unsubstituted C 3 -C 8 cycloalkylene, substituted or unsubstituted 3 to 8 membered heterocycloalkylene, substituted or unsubstituted C 6 -C 10 arylene, or substituted or unsubstituted 5 to 10 membered heteroarylene.

9. The nucleotide analogue of claim 1 , wherein L 4 is a bond or an unsubstituted C 1 -C 4 alkylene.

10. The nucleotide analogue of claim 1 , wherein R 4 is substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted 2 to 6 membered heteroalkyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, substituted or unsubstituted 3 to 6 membered heterocycloalkyl, substituted or unsubstituted phenyl, or substituted or unsubstituted 5 to 6 membered heteroaryl.

11. The nucleotide analogue of claim 1 , wherein R 4 is substituted C 1 -C 6 alkyl.

12. The nucleotide analogue of claim 1 , wherein R 4 is unsubstituted C 1 -C 6 alkyl.

13. The nucleotide analogue of claim 1 , wherein R 4 is substituted phenyl.

14. The nucleotide analogue of claim 1 , wherein R 4 is unsubstituted phenyl.

15. The nucleotide analogue of claim 1 , wherein R 5 is a detectable label.

16. A nucleic acid polymerase complex, wherein said nucleic acid polymerase is bound to a nucleotide analogue of claim 1 .

17. A method for sequencing a nucleic acid, comprising:

(i) incorporating in series with a nucleic acid polymerase, within a reaction vessel, one of four different labeled nucleotide analogues into a primer to create an extension strand, wherein said primer is hybridized to said nucleic acid and wherein each of the four different labeled nucleotide analogues comprise a unique detectable label;

(ii) detecting said unique detectable label of each incorporated nucleotide analogue, so as to thereby identify each incorporated nucleotide analogue in said extension strand, thereby sequencing the nucleic acid;

wherein each of said four different labeled nucleotide analogues is of the structure of claim 1 .

18. A method of incorporating a nucleotide analogue into a primer, the method comprising combining a polymerase, a primer hybridized to nucleic acid template and a nucleotide analogue within a reaction vessel and allowing said polymerase to incorporate said nucleotide analogue into said primer thereby forming an extended primer, wherein said nucleotide analogue is of the structure of claim 1 .

Assignments (1)

Continuity (2)

References Cited (206)

Cited By (1)

US 12,577,271