Method for labeling of aldehyde containing target molecules

Patent metadata

Field	Value
Publication number	US-11293060-B2
Application number	US-201916596539-A
Country	US
Kind code	B2
Filing date	Oct 8, 2019
Priority date	Apr 12, 2017
Publication date	Apr 5, 2022
Grant date	Apr 5, 2022

Abstract

The present invention relates to improving the processing rate of a sequencing reaction, for example in a nanopore sequencing reaction, by means of using improved nucleoside-tags. The tags are linked to the nucleoside phosphate via a Pictet Spengler reaction. Exemplary sequencing reactions that are improved by the present methods include nanopore-based nucleic acid sequencing-by-synthesis reactions.

First claim

The invention claimed is:

1. A method of sequencing a nucleic acid using a tagged nucleoside, wherein the tag has been linked to the nucleoside phosphate via a Pictet Spengler reaction, and the tagged nucleoside has the formula:

wherein the method comprises (i) adding the tagged nucleoside to a reaction chamber; (ii) carrying out a polymerization reaction with the aid of a polymerase, therby incorporating the tagged nucleoside into a complement of the nucleic acid, and (iii) detecting the identity of the tag by altered resistance, current, voltage, or capacitance.

2. The method according to claim 1 , wherein the tag comprises a terminal N-(2-aminoethyl) pyrrole moiety has been linked to an aldehyde moiety connected via a linker to the 5′-end of a nucleoside phosphate.

3. The method according to claim 2 , wherein the nucleoside phosphate is a nucleoside hexaphosphate.

4. The method according to claim 1 , wherein the sequencing reaction is a sequencing by synthesis reaction.

5. The method according to claim 1 , wherein the sequencing reaction is a single molecule sequencing reaction.

6. The method according to claim 5 , wherein the sequencing reaction is a nanopore sequencing reaction.

7. The method according to claim 5 , wherein the tag is a fluorescent tag.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Bergmann Frank
• Chau Ian Quang
• Crisalli Peter
• Kallewaard-Lum Hannah Marie
• Kuchelmeister Hannes
• Pomplun Sebastian Johannes
• Saqib Hasham
• Wellner Christian

Classifications

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• C07D519/00

  Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00 · CPC title

• C07H21/02

  with ribosyl as saccharide radical · CPC title

• G01N33/76

  Human chorionic gonadotropin {including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors} · CPC title