# US 2014/0154688 A1 · Reagent container for amplifying nucleic acid, method of preparing the reagent container, method of storing the reagent, and microfluidic system for nucleic acid analysis

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- Number: US 2014/0154688 A1
- Document ID: US20140154688A1
- Status: Published application
- Kind: A1
- Application number: 14/095,784
- Filed: 2013-12-03
- Published: 2014-06-05
- Earliest claimed: 2012-12-03
- Assignees: Samsung Electronics Co., Ltd.
- Applicants: Samsung Electronics Co. Ltd.
- Inventors: Sung-ouk JUNG; Sung-min Chi; Soo-kwan Lee; Sung-hong Kwon; Joon-ho Kim; Kak Namkoong; Chin-sung Park; Kyu-youn Hwang
- Web page: https://patentgrep.com/patent/US20140154688A1

## CPC

- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- 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 3/527: {for a plurality of reagents}
- B01L 3/502707: {characterised by the manufacture of the container or its components}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- B01L 2200/04: Exchange or ejection of cartridges, containers or reservoirs
- B01L 2200/0684: Venting, avoiding backpressure, avoid gas bubbles
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2200/141: Preventing contamination, tampering
- B01L 2200/16: Reagents, handling or storing thereof
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0861: Configuration of multiple channels and/or chambers in a single devices
- B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
- B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution
- B01L 2400/0487: fluid pressure, pneumatics
- 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
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

Provided is a reagent container that can maintain stability of a reagent for a long period of time, a method of preparing the reagent container, a method of storing the reagent, and microfluidic systems for conducting cell binding, lysis, nucleic acid extraction, and amplification.

## Claims

### Claim 1

A kit comprising
a reagent container comprising a first well and a second well, wherein
the first well contains a first reagent comprising a nucleotide, a nucleic acid component, an enzyme, or a combination thereof without a reaction buffer; and
the second well contains a second reagent comprising a reaction buffer for amplifying a nucleic acid.

### Claim 2 (depends on 1)

The kit of claim 1, wherein the first reagent comprises nucleotide is a deoxynucleotide triphosphate, a ribonucleotide triphosphate, or combination thereof; a nucleic acid primer; and, optionally, a probe nucleic acid.

### Claim 3 (depends on 1)

The kit of claim 1, wherein the first reagent further comprises an enzyme or a stabilizer.

### Claim 4 (depends on 1)

The kit of claim 1, wherein the first reagent is solidified.

### Claim 5 (depends on 1)

The kit of claim 1, wherein the second reagent is solidified.

### Claim 6 (depends on 1)

The kit of claim 1, wherein the first reagent is lyophilized.

### Claim 7 (depends on 1)

The kit of claim 1, wherein the second reagent is lyophilized.

### Claim 8 (depends on 1)

The kit of claim 1, wherein the reagent container further comprises a connection part that connects the first well and the second well to each other.

### Claim 9 (depends on 8)

The kit of claim 8, wherein the connection part is a groove, a channel, a partition, or a film.

### Claim 10 (depends on 1)

The kit of claim 1, wherein the container comprises a plurality of protrusions, and the first well and the second well are provided by two subgrooves separated from each other and recessed in a predetermined shape on the plurality of protrusions.

### Claim 11 (depends on 10)

The kit of claim 10, wherein the wells have a curved shape comprising two curved side portions that meet at the center of the well, wherein the center of the well is narrower than the sides of the well.

### Claim 12 (depends on 10)

The kit of claim 10, wherein an external angle 8 formed by corners of both sides of the first well and the second well at a position having the narrowest width is in a range of about 30 degrees to about 90 degrees.

### Claim 13 (depends on 1)

The kit of claim 1, wherein the first well comprises a first aperture, and the second well comprises a second aperture, wherein the aperture is provided by an open groove.

### Claim 14 (depends on 13)

The kit of claim 13, wherein the reagent container is attached to a rehydration chamber, and the open groove of the first aperture and the open groove of the second aperture contact an outer surface of the rehydration chamber to form one channel.

### Claim 15 (depends on 1)

A microfluidic system for analyzing nucleic acid, the microfluidic system comprising:
a rehydration chamber;
the kit of claim 1 attached to the rehydration chamber;
an amplification chamber; and
a flow channel system providing a fluid flow path between the rehydration chamber and the amplification chamber.

### Claim 16 (depends on 15)

The microfluidic system of claim 15, wherein the rehydration chamber comprises two separated sub-chambers.

### Claim 17 (depends on 16)

The microfluidic system of claim 16, wherein a side of the sub-chamber has a curved shape and a width of a flow path of the introduced cell lysate is smallest at a center portion thereof.

### Claim 18 (depends on 15)

The microfluidic system of claim 15, further comprising:
a reagent supply device including a sample chamber in which a sample as an examination target is injected, one or more reagent chambers in which a reagent for extracting nucleic acid from the sample is injected, and a waste chamber in which the used reagent is discarded;
a binding-lysis chamber in which cells are captured from the sample, the captured cells are lysed to form a cell lysate containing nucleic acid, and a plurality of particles for cell binding are disposed; and
a flow channel system having an outlet and a plurality of inlets connected to the reagent supply device and forming an integrated fluid flow between the binding-lysis chamber, the rehydration chambers, and the amplification chambers.

### Claim 19

A method of storing a reagent, the method comprising:
disposing a first reagent comprising a nucleotide or nucleic acid into a first well of a reagent container, wherein the first well of the reagent container does not contain a reaction buffer;
disposing a second reagent comprising a reaction buffer into a second well of a reagent container; and
solidifying the first reagent by drying;
wherein the wherein the first and the second reagent are for amplifying nucleic acids.

### Claim 20 (depends on 19)

The method of claim 19, wherein the first reagent further comprises an enzyme.