REVERSIBLE TERMINATOR NUCLEOTIDES AND METHODS OF USE

• US 20080050780A1
• Filed: 06/14/2007
• Published: 02/28/2008
• Est. Priority Date: 06/30/2006
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: LEE, Linda, Zon, Gerald
• Granted Patent: US 7,553,949 B2
• Time in Patent Office: Days
• US Class Current: 435/91.1
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/525 Phosphatase
• CPC Class Codes: C12Q 2525/101 incorporating non-naturally occurring nucleotides, e.g. inosine

Abstract

Disclosed herein a reversible terminator nucleotides and methods of use.

Claims

1. A nucleoside comprising a base and a sugar in which a label is attached to said base and a protecting group is attached to said sugar, wherein said label and said protecting group can be detached from said nucleoside by an enzyme.

2. The nucleoside of claim 1, wherein said label and said protecting group can be detached from said nucleoside under identical conditions.

3. The nucleoside of claim 1, wherein said label and said protecting group comprise a phosphate moiety.

4. The nucleoside of claim 1, wherein said label and said protecting group comprise a SO₃ moiety.

5. The nucleoside of claim 1, wherein said label and said protecting group comprise a C(O)R moiety.

6-14. -14. (canceled)

15. A nucleotide comprising a base and a sugar in which a label is attached to said base and a protecting group is attached to said sugar, wherein said label and said protecting group can be detached from said nucleotide by an enzyme.

16. The nucleotide of claim 15, wherein said label and said protecting group can be detached from said nucleoside under identical conditions.

17. The nucleotide of claim 15, wherein said label and said protecting group comprise a phosphate moiety.

18. The nucleotide of claim 15, wherein said label and said protecting group comprise a SO₃ moiety.

19. The nucleotide of claim 15, wherein said label and said protecting group comprise a C(O)R moiety.

20. The nucleotide of claim 15, wherein said enzyme is a phosphatase.

21. The nucleotide of claim 20, wherein said phosphatase is shrimp alkaline phosphatase.

43. A method of extending a primer nucleic acid comprising:

contacting a primer hybridized to a template polynucleotide with a polymerase and the nucleotide of claim 15 under conditions suitable for said polymerase to extend said primer in a template dependent manner by the addition of said nucleotide to the 3′

terminus of said primer.

22-42. -42. (canceled)

44-49. -49. (canceled)

50. (canceled)