Methods of functionalizing magnetic particles and methods of generating amplicons using magnetic particles

Patent metadata

Field	Value
Publication number	US-2025011834-A1
Application number	US-202418753740-A
Country	US
Kind code	A1
Filing date	Jun 25, 2024
Priority date	Jun 29, 2023
Publication date	Jan 9, 2025
Grant date	—

Abstract

This application relates to functionalized magnetic particles and methods of using magnetic particles to generate amplicons. In some examples, a method of modifying a magnetic particle comprising a first functional group includes contacting the magnetic particle with a first molecule comprising a polymer coupled to second and third functional groups, wherein the first functional group reacts with the second functional group to form a bond via which the polymer is coupled to the magnetic particle. The method may include contacting the magnetic particle with a second molecule, the second molecule comprising an oligonucleotide coupled to a fourth functional group, wherein the third functional group reacts with the fourth functional group to form a bond via which the oligonucleotide is coupled to the magnetic particle.

First claim

1 . A method of modifying a magnetic particle comprising a first functional group, the method comprising:
contacting the magnetic particle with a first molecule comprising a polymer coupled to second and third functional groups, wherein the first functional group reacts with the second functional group to form a bond via which the polymer is coupled to the magnetic particle; and contacting the magnetic particle with a second molecule, the second molecule comprising an oligonucleotide coupled to a fourth functional group, wherein the third functional group reacts with the fourth functional group to form a bond via which the oligonucleotide is coupled to the magnetic particle.

2 . The method of claim 1 , wherein the oligonucleotide comprises an amplification primer.

3 . The method of claim 1 , wherein the second and third functional groups are of the same type as one another.

4 . The method of claim 1 , wherein the second and third functional groups are of different types than one another.

5 . The method of claim 1 , wherein the polymer comprises a hydrogel.

6 . The method of claim 5 , wherein the hydrogel comprises polyacrylamide.

7 . The method of claim 1 , wherein the magnetic particle has a diameter of about 100 nm to about 300 nm.

8 . A method of modifying a magnetic particle coupled to a polymer comprising a first functional group, the method comprising:
contacting the magnetic particle with a molecule comprising an oligonucleotide coupled to a second functional group, wherein the second functional group reacts with the first functional group to form a bond via which the oligonucleotide is coupled to the magnetic particle.

9 . A composition made by the method of claim 1 .

10 . A composition, comprising:
a magnetic particle; a polymer coupled to the magnetic particle via a reaction product of a first functional group coupled to the magnetic particle and a second functional group coupled to the polymer; and a plurality of oligonucleotides coupled to the polymer via reaction products of third functional groups coupled to the polymer and fourth functional groups coupled to the oligonucleotides.

11 . A method of using the composition of claim 10 , the method comprising:
hybridizing a template polynucleotide to a first oligonucleotide of the plurality; and amplifying the hybridized template polynucleotide using additional oligonucleotides of the plurality to generate a plurality of amplicons coupled to the polymer via reaction products of the third and fourth functional groups.

12 . The method of claim 11 , further comprising, after the amplifying, disposing the magnetic particle within a flow cell.

13 . The method of claim 12 , wherein disposing the magnetic particle within the flow cell comprises electrostatically attracting the magnetic particle to a region of the flow cell.

14 . The method of claim 11 , further comprising, before the amplifying, disposing the magnetic particle within a flow cell.

15 . A device comprising:
a flow cell; and the composition of claim 10 located within the flow cell.

16 . The device of claim 15 , wherein the flow cell comprises a positively charged surface to which the composition is noncovalently bound through an electrostatic force.

17 . A composition, comprising:
a magnetic particle; and a molecule coupled to the magnetic particle, the molecule comprising:
a polymer coupled to the magnetic particle via a reaction product of first and second functional groups; and
a plurality of polynucleotide amplicons coupled to the polymer via reaction products of third and fourth functional groups.

18 . A device comprising:
a flow cell; and the composition of claim 17 located within the flow cell.

19 . The device of claim 18 , wherein the flow cell comprises a positively charged surface to which the composition is noncovalently bound through an electrostatic force.

20 . A method of using the device of claim 19 , the method comprising:
sequencing the plurality of polynucleotide amplicons within the flow cell.

21 - 35 . (canceled)

Assignees

• Illumina Inc

Inventors

• Gal Teodora
• Von Hatten Xavier
• Artioli Gianluca
• Tovey Will
• Nguyen Nam
• La Rosa Angelo

Classifications

• C12Q1/6876

  Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• B01J19/0046

  Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays (synthesis methods per se C40B50/00) · CPC title

• C12Q1/6846

  Common amplification features · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title