# US 12,109,562 B2 · Nucleic acid extraction apparatus and method of use thereof

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- Number: US 12,109,562 B2
- Document ID: US12109562B2
- Status: Granted
- Kind: B2
- Application number: 17/882,382
- Filed: 2022-08-05
- Published: 2022-11-24
- Granted: 2024-10-08
- Earliest claimed: 2018-11-26
- Assignees: Hangzhou Bigfish Bio-Tech Co., Ltd.
- Applicants: Hangzhou Bigfish Bio-Tech Co., Ltd.
- Inventors: Lianyi Xie
- Web page: https://patentgrep.com/patent/US12109562B2

## CPC

- B01L 3/502: {with fluid transport, e.g. in multi-compartment structures}
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- B01L 3/502723: {characterised by venting arrangements}
- C12N 15/1013: {by using magnetic beads}
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- C12Q 1/6846: {Common amplification features}
- C12Q 1/6848: characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
- B01L 2200/0621: Control of the sequence of chambers filled or emptied
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2200/16: Reagents, handling or storing thereof
- B01L 2300/0803: Disc shape
- B01L 2300/0861: Configuration of multiple channels and/or chambers in a single devices
- B01L 2300/087: Multiple sequential chambers
- B01L 2400/0478: pistons
- C12Q 1/686: Polymerase chain reaction [PCR]
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention discloses a nucleic acid extraction apparatus and a method using the apparatus for extracting nucleic acids and amplifying target nucleic acids. The nucleic acid extraction apparatus comprising: a nucleic acid extraction element, a waste storage chamber, and a reaction chamber, wherein the reaction chamber is selectively in communication with the nucleic acid extraction element and the waste storage chamber, and the apparatus realizes fluid exchange through the pressure between the waste storage chamber and the reaction chamber. In the present invention, by using an integrated control system, the conventional manual process of nucleic acid extraction and purification is integrated into a fully automatic closed process, making the operation more conveniently and quickly and improving the efficiency of experimental work.

## Claims

### Claim 1

Nucleic acid extraction apparatus, comprising:
a plurality of nucleic acid extraction chambers;
a reaction chamber, wherein the reaction chamber is selectively in communication with one of the nucleic acid extraction chambers;
a hollow piston disposed in the reaction chamber and configured to move between a first position and a second position in the reaction chamber to change a pressure in the reaction chamber;
a magnetic bead disposed in the reaction chamber for binding a nucleic acid;
a receiving chamber for accommodating a permanent magnet as to fix the magnetic bead on an outer side wall of the receiving chamber;
a nucleic acid amplification element, wherein the nucleic acid amplification element is a nucleic acid amplification chamber that is in fluidic communication with the reaction chamber;
wherein the receiving chamber is disposed in the hollow piston, and the hollow piston is movable relative to the receiving chamber; and wherein a recess is provided on the outer side wall at a bottom of the hollow piston, and when the hollow piston is at the second position, the recess and the outer side wall of the receiving chamber form a space for accommodating the magnetic bead therein.

### Claim 2 (depends on 1)

The apparatus according to claim 1, wherein the nucleic acid amplification chamber is a PCR reaction tube.

### Claim 3 (depends on 1)

The apparatus according to claim 1, wherein the nucleic acid extraction chambers comprise a lysis chamber with lysate reagent for receiving a sample; a wash chamber for adding and storing washing liquid; an eluent chamber for adding and storing eluent reagent.

### Claim 4 (depends on 3)

The apparatus according to claim 3, wherein the reaction chamber is selectively in communication with one or more of the lysis chamber, the wash chamber or the eluent chamber of the nucleic acid extraction chambers.

### Claim 5 (depends on 4)

The apparatus according to claim 4, wherein the apparatus further comprises a microfluidic channel for fluidic communication with the reaction chamber; the lysis chamber, the wash chamber or the eluent chamber of the nucleic acid extraction chambers.

### Claim 6 (depends on 5)

The apparatus according to claim 5, wherein the microfluidic channel is radially extended on a rotary disk, and the reaction chamber is in communication with or not in communication with one or more of the lysis chamber, the wash chamber or the eluent chamber of the nucleic acid extraction chambers by rotation of the rotary disk.

### Claim 7 (depends on 1)

The apparatus according to claim 1, wherein the apparatus further comprises a rotary disk, the reaction chamber is in communication with or not in communication with one or more of the nucleic acid extraction chambers by rotation of the rotary disk.

### Claim 8 (depends on 7)

The apparatus according to claim 7, wherein a microfluidic channel is disposed in the rotary disk.

### Claim 9 (depends on 8)

The apparatus according to claim 8, further comprising a wash chamber, wherein the wash chamber comprises a primary wash chamber, a secondary wash chamber and a tertiary wash chamber, one or more of the primary wash chamber, the secondary wash chamber or the tertiary wash chamber being in fluidic communication with the reaction chamber, respectively.

### Claim 10 (depends on 1)

The apparatus according to claim 1, wherein a rotary disk disposed at a bottom of the shell, wherein the rotary disk includes a microfluidic channel for fluidic communication with the reaction chamber and one of the nucleic acid extraction chambers.

### Claim 11 (depends on 10)

The apparatus according to claim 10, wherein a fluid in the one of the nucleic acid extraction chambers enters into the reaction chamber by a movement of the hollow piston.

### Claim 12 (depends on 11)

The apparatus according to claim 11, wherein the movement of the hollow piston is an upward movement along the receiving chamber.

### Claim 13 (depends on 1)

The apparatus according to claim 1, further comprising a wash chamber, wherein the wash chamber comprises a primary wash chamber, a secondary wash chamber and a tertiary wash chamber, one or more of the primary wash chamber, the secondary wash chamber or the tertiary wash chamber being in fluidic communication with the reaction chamber.

### Claim 14 (depends on 13)

The apparatus according to claim 13, further comprising a waste chamber and a lysis chamber, wherein the waste chamber is not in fluidic communication with the reaction chamber when the reaction chamber is in fluidic communication with the lysis chamber and the reaction chamber is not in fluidic communication with the lysis chamber when the waste chamber is in fluidic communication with the reaction chamber.

### Claim 15 (depends on 14)

The apparatus according to claim 14, wherein a fluid in the lysis chamber enters the reaction chamber by a movement of the piston along the receiving chamber.

### Claim 16 (depends on 1)

The apparatus according to claim 1, wherein the apparatus further comprises a nucleic acid amplification element comprising a PCR reaction tube that is in fluidic communication with a PCR reaction solution chamber.

### Claim 17 (depends on 16)

The apparatus according to claim 16, wherein a sidewall of the PCR reaction solution chamber is provided with an input port and a discharge port that are in fluidic communication with the PCR reaction tube.

### Claim 18 (depends on 17)

The apparatus according to claim 17, wherein the PCR reaction solution chamber comprises a primary PCR reaction solution chamber and a secondary PCR reaction solution chamber, wherein the primary PCR reaction solution chamber is in fluidic communication with the reaction chamber, wherein an input port is disposed at a bottom of the sidewall of the primary PCR reaction solution chamber, and a discharge port disposed at a top of the sidewall of the secondary PCR reaction solution chamber.