Nucleotide analogs for real-time sequencing method for single molecule

• US 10,087,207 B2
• Filed: 11/25/2016
• Issued: 10/02/2018
• Est. Priority Date: 02/19/2014
• Status: Active Grant

• Current Assignee: Personal Genomics, Inc.
• Original Assignee: Personal Genomics, Inc.
• Inventors: Fu, Tsu-Ju, Chang, Shang-Chia, Fann, Jenn-Yeh
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US15/361,083
• Publication Number:
  US 20170073752A1
• Time in Patent Office: 676 Days
• Field of Search: 435 61, 435 911, 435 912, 536 41
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2525/113 incorporating modified backbone
• CPC Class Codes: C12Q 2563/107 fluorescence

Abstract

This invention is to describe nucleotide analogs used in a method for the determination of a nucleic acid sequence. The method involves an enzyme, an enzyme complex or plural number of enzymes with more than one enzymatic activity and a set of nucleotide analogs, to achieve high signal readout accuracy in nucleic acid sequencing by making each signal to have a long signaling time span which allows a higher signal clarity.

Claims

1. A nucleotide analog comprising a base moiety and at least one label moiety comprising a photo-detectable label, wherein the photo-detectable label is connected to a 3′

position of a sugar of the nucleotide analog via a coupler group, wherein the coupler group is ester, and the at least one label moiety includes a linker group selected from alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, polyethylene glycol, ester, sulfonyl or a combination thereof, connecting the photo-detectable label to the coupler group, and the nucleotide analog has a structure as shown in Formula I or Formula II;

2. The nucleotide analog of claim 1, wherein the photo-detectable label is a fluorophore or a fluorescent dye.

3. The nucleotide analog of claim 1, wherein the nucleotide analog has 3-12 phosphate groups at the 5′

-end of the nucleotide analog.

4. A nucleotide analog comprising a base moiety and at least one label moiety comprising a photo-detectable label, wherein the photo-detectable label is connected to a 3′

position of a sugar of the nucleotide analog via a coupler group, wherein the coupler group is ester, and the at least one label moiety includes a linker group selected from alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, polyethylene glycol, ester, sulfonyl or a combination thereof, connecting the photo-detectable label to the coupler group, and the nucleotide analog has a structure as shown in Formula XVI;

5. The nucleotide analog of claim 4, wherein the photo-detectable label is a fluorophore or a fluorescent dye.

6. The nucleotide analog of claim 4, wherein the nucleotide analog has 3-12 phosphate groups at the 5′

-end of the nucleotide analog.