Nucleotide analogs for sequencing

• US 10,697,009 B2
• Filed: 08/12/2019
• Issued: 06/30/2020
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Ibis Biosciences Incorporated (Abbott Laboratories)
• Original Assignee: Ibis Biosciences Incorporated (Abbott Laboratories)
• Inventors: Hayden, Mark A., Huff, Jeffrey, Eshoo, Mark W., Picuri, John
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US16/538,278
• Publication Number:
  US 20190367976A1
• Time in Patent Office: 323 Days
• Field of Search: 536 41, 536 231, 536 266
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2523/319 Photocleavage, photolysis, photoactivation
• CPC Class Codes: C12Q 2525/117 incorporating modified base
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2563/116 electrical properties of nucleic acids, e.g. impedance, conductivity or resistance
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label

Abstract

Provided herein is technology relating to sequencing nucleic acids, but not exclusively, to compositions, methods, systems, and kits related to nucleotides comprising an electrochemically detectable moiety and one or more photolabile synthesis-inhibiting moieties.

Claims

1. A system for sequencing a nucleic acid, comprising:

a) a first nucleotide analog comprising an adenine nucleotide;

b) a second nucleotide analog comprising a cytosine nucleotide;

c) a third nucleotide analog comprising a guanine nucleotide; and

d) a fourth nucleotide analog comprising a thymine or a uracil nucleotide;

wherein one or more of said first nucleotide analog, said second nucleotide analog, said third nucleotide analog and said fourth nucleotide analog comprises;

i) a phosphate moiety;

ii) a base;

iii) a sugar;

iv) a first photocleavable terminating moiety attached to said nucleotide analog by a first photocleavable linker, wherein said photocleavable terminating moiety is electrochemically detectable by accepting or donating at least one electron during an electrochemical reaction; and

v) a second photocleavable terminating moiety attached to said nucleotide analog by a second photocleavable linker.

2. The system of claim 1, wherein said first nucleotide analog comprises a first electrochemically detectable moiety or a first fluorescently detectable moiety, said second nucleotide analog comprises a second electrochemically detectable moiety or a second fluorescently detectable moiety, said third nucleotide analog comprises a third electrochemically detectable moiety or a third fluorescently detectable moiety, and said fourth nucleotide analog comprises a fourth electrochemically detectable moiety or a fourth fluorescently detectable moiety.

3. The system of claim 1, wherein said electrochemically detectable, photocleavable terminating moiety is attached to said phosphate moiety, said base or said sugar of said nucleotide analog.

4. The system of claim 1, wherein said phosphate moiety is a triphosphate moiety, a diphosphate moiety or a monophosphate moiety.

5. The system of claim 1, wherein said electrochemically detectable, photocleavable terminating moiety is a group imparting polymerase termination properties to said nucleotide analog.

6. The system of claim 1, further comprising a substrate comprising a zero mode waveguide.

7. The system of claim 1, further comprising an electrical detection element and/or a fluorescence detection element.

8. The system of claim 1, further comprising a polymerase.

9. The system of claim 1, further comprising an illumination source.

10. The system of claim 1, further comprising a nucleic acid sequencer.

11. The system of claim 1, further comprising a sample sequence data storage.

12. The system of claim 1, further comprising a reference sequence data storage.

13. The system of claim 1, further comprising an analytics computing device, server or node.

14. The system of claim 1, further comprising a software component.

15. The system of claim 1, further comprising a sample processing unit.

16. The system of claim 1, further comprising a reagent delivery system.

17. The system of claim 1, further comprising a reagent reservoir.

18. The system of claim 1, further comprising a manipulator, a stepper motor and/or a pump.