Nucleic acid synthesis and purification device, use thereof, and nucleic acid synthesis and purification method

Patent metadata

Field	Value
Publication number	US-12247198-B2
Application number	US-201817311697-A
Country	US
Kind code	B2
Filing date	Dec 14, 2018
Priority date	Dec 14, 2018
Publication date	Mar 11, 2025
Grant date	Mar 11, 2025

Abstract

A nucleic acid synthesis device and a nucleic acid purification device, uses thereof, and a nucleic acid synthesis method and a nucleic acid purification method. The nucleic acid synthesis device includes a solid support, and the solid support includes a controlled pore glass (CPG), the CPG is an unmodified and bare CPG, a surface of the CPG has a hydroxyl group, and the hydroxyl group is attachable, though a covalent bond, to a phosphoramidite-protected nucleotide monomer or multimer for synthesis of nucleic acid. The nucleic acid synthesis device of the present disclosure can be used for not only synthesis of an oligonucleotide primer, but also for purification of enzymatic digestion and PCR product by using the oligonucleotide primer immobilized on the CPG, and has advantages of simple structure, small volume, light weight, high efficiency, low costs, and diversified functions.

First claim

What is claimed is:

1. A nucleic acid purification method, comprising:
adding a solution containing a nucleic acid to be purified into a nucleic acid purification device for incubation, the nucleic acid purification device comprising a solid support, wherein the solid support comprises a controlled pore glass (CPG), the CPG is an unmodified and bare CPG, a surface of the CPG has a hydroxyl group, and the hydroxyl group is covalently bonded directly to an oligonucleotide sequence, and wherein the nucleic acid to be purified is complementary hybridized or randomly hybridized with all of or a part of the fragment of oligonucleotide sequence in the nucleic acid purification device;
eluting the nucleic acids to be purified from the nucleic acid purification device.

2. The nucleic acid purification method according to claim 1 , wherein the fragment of oligonucleotide sequence is specifically complementary hybridized or randomly hybridized with all of or part of a regional fragment at a 3′-end or/and 5′-end of the nucleic acid to be purified.

3. The nucleic acid purification method according to claim 1 , wherein the nucleic acid to be purified is DNA, and the nucleic acid purification device is a DNA purification device.

4. The nucleic acid purification method according to claim 1 , further comprising a screen plate, wherein the screen plate comprises an upper screen plate and a lower screen plate, and the CPG is disposed between the upper screen plate and the lower screen plate.

5. The nucleic acid purification method according to claim 4 , wherein the upper screen plate is hydrophilically treated or selected from hydrophilic materials, and the lower screen plate is selected from hydrophobic materials.

6. The nucleic acid purification method according to claim 1 , wherein the solid support further comprises a thermoplastic polymer resin, and the thermoplastic polymer resin is sintered with the CPG to form a Controlled Pore Glass Frit (CPG Frit).

7. The nucleic acid purification method according to claim 1 , wherein the nucleic acid purification device further comprises a receiving device, and wherein the solid support is disposed in the receiving device.

8. The nucleic acid purification method according to claim 7 , wherein the receiving device is a hollow column tube.

9. The nucleic acid purification method according to claim 8 , wherein at least one assembly site is provided within the hollow column tube, the assembly site defines a cylinder, the solid support is in a shape of a cylinder, the solid support tangentially fits the assembly site, and a height of the assembly site is greater than a thickness of the solid support.

10. The nucleic acid purification method according to claim 1 , wherein the nucleic acid to be purified is a single-stranded nucleic acid.

11. The nucleic acid purification method according to claim 1 , wherein the nucleic acid purification device is subjected to an ammonolysis deprotection treatment prior to the incubation with the solution.

12. The nucleic acid purification method according to claim 1 , wherein the incubation is followed by washing with a mixture of ethanol and water as a mobile phase to remove unbound nucleic acid and/or enzyme and/or buffer, and eluting the nucleic acids to be purified from the nucleic acid purification device.

Assignees

• Bgi Shenzhen

Inventors

• Fang Xin
• Huang Xiaoluo
• Shen Yue
• Xu Xun

Classifications

• B01J2219/00759

  Purification of compounds synthesised · CPC title

• B01J2219/00497

  Features relating to the solid phase supports · 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

• C12M1/40

  Apparatus specially designed for the use of free, immobilised, or carrier-bound enzymes, e.g. apparatus containing a fluidised bed of immobilised enzymes · CPC title

• C12M1/00

  Apparatus for enzymology or microbiology · CPC title