KR101771493B1 - A micro polymerase chain reaction chip and the manufacturing method thereof - Google Patents
A micro polymerase chain reaction chip and the manufacturing method thereof Download PDFInfo
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- KR101771493B1 KR101771493B1 KR1020150174831A KR20150174831A KR101771493B1 KR 101771493 B1 KR101771493 B1 KR 101771493B1 KR 1020150174831 A KR1020150174831 A KR 1020150174831A KR 20150174831 A KR20150174831 A KR 20150174831A KR 101771493 B1 KR101771493 B1 KR 101771493B1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
- B01L3/50851—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates specially adapted for heating or cooling samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0627—Sensor or part of a sensor is integrated
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/16—Surface properties and coatings
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
- B01L2400/0633—Valves, specific forms thereof with moving parts
- B01L2400/0644—Valves, specific forms thereof with moving parts rotary valves
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Abstract
Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in an effort to solve the problems of the prior art described above, and its object is to minimize the bubble formation, evaporation, and absorption of fluid by performing a paralyn coating inside a reaction chamber in which a fluid exists. In addition, it is possible to move the fluid by deformation caused by applying a force to the injection port and the discharge port by using a polymer having an elastic force, and when the force is not applied, the elasticity returns to the original state and the evaporation of the fluid can be reduced by blocking the injection port and the discharge port , And a polymer chip made of a polymer material having a complicated fabrication process. In order to accomplish the object of the present invention as described above, a micro PCR chip according to an embodiment of the present invention is disclosed. The PCR chip may be made of a polymer material, and the inside of the reaction chamber in which the fluid exists may be paralin-coated. The reaction chamber may include an inlet and an outlet.
Description
The present invention relates to a micro-polymerase chain reaction chip and a method of manufacturing the same, and more particularly, to a cuff-shaped neural electrode capable of taking advantage of different materials by making materials of a structure and a window different from each other, and a manufacturing method thereof will be.
Polymerase chain reaction (PCR) is a method of amplifying a specific genetic material that is desired to be detected. When a fluid containing DNA is subjected to an amplification process, bubble formation, evaporation, and absorption occur. Among them, the bubble formation causes the temperature to be unbalanced and the temperature difference in the fluid is generated. In addition, evaporation and absorption may interfere with the mixing of the buffer solution and the primer, resulting in poor DNA amplification. In the case of viral DNA, secondary infection due to evaporation may occur. To reduce these losses, various materials such as oil, oxide film, and paralin-C were used as coating materials for chips made of PC (Polycarbonate), PDMS (polydimethylsiloxane), and silicon. Among them, Paralyn-C is a frequently used material for coating fluidic chips and has low permeability for safety and moisture.
The PCR chip according to the prior art is a PDMS chip, which prevents vaporization of the fluid through the external Parallel-C coating. However, due to the structural problem that the fluid is absorbed into the chip and the inlet and outlet are open, exist.
When the fluid is injected through the injection port and the internal air is discharged through the discharge port, a structure having both the injection port and the discharge port can be used.
When the fluid is injected into the injection port of the chip, the internal pressure is increased. At the same time, the air inside is discharged through the discharge port. On the other hand, the fluid injected into the chip through the injection port can not prevent evaporation because the injection port and the discharge port are open.
Digital PCR has been developed to improve the sensitivity and precision of existing PCR technology. It is a technique to separate PCR samples in multiple compartments and then perform reactions individually in each compartment. To compartmentalize the PCR sample, the prior art (Figure 1) used a method in which the PCR sample was injected using vacuum pressure and the oil was injected directly using a syringe. However, it is inconvenient to keep the inside of the fluid channel in a vacuum state and repeatedly use a general tape for sealing at the time of PCR amplification. Also, due to the leakage generated when the tape is removed and the fluid is inserted, the positive pressure enters the fluid channel, making it difficult to efficiently load the sample. In addition, after the PCR sample is injected with vacuum pressure, there is a further inconvenience to inject the oil directly for the chamber compartment.
Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in an effort to solve the problems of the prior art described above, and its object is to minimize the bubble formation, evaporation, and absorption of fluid by performing a paralyn coating inside a reaction chamber in which a fluid exists.
In addition, when the force is applied to the injection port and the discharge port using the elastic polymer, the fluid can be moved. When the force is not applied, the elasticity can return to the original state and the evaporation of the fluid can be reduced by blocking the injection port and the discharge port. The present invention aims at solving the problems of the prior art by proposing a polymer chip-type PCR chip having a complicated manufacturing process.
In order to accomplish the object of the present invention as described above, a micro PCR chip according to an embodiment of the present invention is disclosed.
The PCR chip may be made of a polymer material, and the inside of the reaction chamber in which the fluid exists may be paralin-coated.
The reaction chamber may include an inlet and an outlet.
When the fluid is injected into the reaction chamber through the injection port, deformation may occur in the injection port and the discharge port, and the air inside the discharge port may be discharged due to the pressure.
If there is no injection of fluid through the injection port, the discharge port can be closed by the elastic force of the polymer material.
The PCR chip may include a rotary valve.
The rotary valve may include the inlet, the outlet and the loading chamber.
The PCR sample and oil may be placed into the loading chamber and the PCR sample and oil may be injected into the reaction chamber by rotating the rotatable valve to align the loading chamber with the input channel or output channel.
When the PCR sample and the oil are injected into the reaction chamber, the rotary valve may be rotated to close the input channel or the output channel.
The inlet structure of the reaction chamber may be in the form of a diffuser nozzle.
The parallax may be coated by fabricating the PDMS chip in a mold, coating the PDMS with a paralin layer to fill the holes of the PDMS, and removing the paralin layer.
According to the structure of the present invention as described above, the bubble formation, evaporation and absorption of fluid can be minimized, and a PCR chip having a complicated manufacturing process can be manufactured.
Those skilled in the art will appreciate that the effects derived from the present invention are not limited to the above description and can be widely accepted.
Figure 1 shows a PCR chip according to the prior art.
2 shows a paralin coating method according to an embodiment of the present invention.
Fig. 3 shows the change of the fluid in the reaction chamber of the PCR chip with or without the paralin coating.
4 illustrates the operation principle of a reaction chamber of a PCR chip according to an embodiment of the present invention.
5 shows a rotary valve of a PCR chip according to an embodiment of the present invention.
Specific structural and functional descriptions of embodiments of the invention disclosed herein are merely illustrative for purposes of illustrating embodiments of the invention and that the embodiments according to the invention may be embodied in various forms, And should not be interpreted as being limited to the embodiments described in the application.
Since the embodiments according to the present invention can make various changes and have various forms, specific embodiments are illustrated in the drawings and described in detail herein. It should be understood, however, that the embodiments according to the concepts of the present invention are not intended to be limited to any particular mode of disclosure, but rather all variations, equivalents, and alternatives falling within the spirit and scope of the present invention.
The terms first and / or second, primary and / or secondary, etc. may be used to describe various components, but the components should not be limited by these terms. The terms are intended to distinguish one element from another, for example, without departing from the scope of the invention in accordance with the concepts of the present invention, the first element may be termed the second element, The second component may also be referred to as a first component.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise," "include," "have," and the like, specify that there is a specified feature, number, step, operation, component, section, element, Steps, operations, elements, parts, or combinations thereof, whether or not explicitly described or implied by the accompanying claims.
Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be construed as meaning consistent with meaning in the context of the relevant art and are not to be construed as ideal or overly formal in meaning unless expressly defined herein .
Hereinafter, a PCR chip according to the present invention will be described in detail with reference to the accompanying drawings.
2 shows a paralin coating method according to an embodiment of the present invention.
2, the parallel-Lin, (a) PDMS produce a chip in a mold and, (b) by coating the parallel-lean layer to PDMS (c) filling the holes in the PDMS, (d) O through the second plasma etching It can be coated by removing the paralin layer. Paralin may be, for example, Paralin-C.
By coating the holes of the PDMS with the paralin coating, it is possible to improve the DNA amplification efficiency by reducing the formation, evaporation, and absorption of the fluid due to holes on the PDMS surface.
Also, by fabricating the structure of the chip using the polymer, evaporation of the injection part and the discharge port can be prevented and the efficiency can be improved.
Fig. 3 shows the change of the fluid in the
When a PCR chip is manufactured, a PCR chip including a
4 shows the operation principle of the
Referring to FIG. 4, when fluid is injected into the
5 shows a rotary valve of a PCR chip according to an embodiment of the present invention.
Referring to FIG. 5, the rotary valve can be applied to a chip for performing digital PCR, so that the vacuum pressure can be efficiently maintained and the operation of the fluid can be simplified. This further prevents evaporation from occurring during PCR amplification.
More specifically, placing the PCR sample and oil in the loading chamber of the rotary valve causes the oil to be placed on top of the PCR sample by the density difference. When the rotary valve is turned so that the input or output channel and the loading chamber are aligned, the PCR sample and oil are automatically injected by vacuum pressure. At this time, since the PCR sample is located below the oil, it is injected first to fill the reaction chamber, and then the incoming oil is filled in the fluid channel, so that each reaction chamber is partitioned. It is also designed to be able to inject oil directly using the inlet / outlet regenerator by turning the rotary valve again to perform the fluid injection more reliably. Finally, when the rotary valve is turned again, the inlet / outlet channel and the loading chamber, or the inlet / outlet regenerative baffle are turned off, thereby blocking the fluid channel.
In the present invention, the structure of the inlet of the reaction chamber to minimize the loss of the PCR sample existing when the oil is injected is adopted as the diffuser nozzle shape. According to this configuration, the PCR sample loss rate is about 3% It is confirmed that the loss ratio is small.
Claims (10)
A reaction chamber in which a fluid is introduced into the reaction chamber, and an injection port communicating with a lower portion of the reaction chamber to allow the fluid to flow into the reaction chamber, And a polymer layer extending in a direction intersecting the extending direction of the reaction chamber and having a discharge port for discharging the internal air of the reaction chamber when the fluid flows into the reaction chamber,
Wherein the polymer layer is deformed to open the injection port and the discharge port when the fluid flows into the channel in accordance with an increase in pressure, and when the input of the fluid into the channel is completed, ≪ / RTI >
Micro PCR chip.
Wherein the inner wall surface of the reaction chamber is a paralin-coated, micro PCR chip.
Wherein the polymer layer is formed of PDMS (polydimethylsiloxane) so that the injection port and the discharge port are deformed depending on whether the fluid is injected into the channel.
Wherein the micro PCR chip comprises a rotatable valve.
Wherein the rotatable valve comprises the inlet, the outlet and the loading chamber.
Placing the PCR sample and the oil in the loading chamber and rotating the rotatable valve to align the loading chamber with the input channel or output channel to inject the PCR sample and oil into the reaction chamber.
And when the PCR sample and the oil are injected into the reaction chamber, the rotary valve is rotated to close the input channel or the output channel.
Wherein the inlet structure of the reaction chamber is in the form of a diffuser nozzle.
The Paralyne,
A micro PCR chip, wherein a PDMS chip is fabricated in a mold, the PDMS is coated with a paralin layer to fill the holes of the PDMS, and the paralin layer is removed.
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KR1020150174831A KR101771493B1 (en) | 2015-12-09 | 2015-12-09 | A micro polymerase chain reaction chip and the manufacturing method thereof |
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KR1020150174831A KR101771493B1 (en) | 2015-12-09 | 2015-12-09 | A micro polymerase chain reaction chip and the manufacturing method thereof |
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KR102115094B1 (en) * | 2018-10-23 | 2020-05-25 | 인제대학교 산학협력단 | Digital polymerase chain reaction (dPCR) system based on microfluidic technology and the method thereof |
CN110452814A (en) * | 2019-07-12 | 2019-11-15 | 北京资和源医疗科技有限公司 | A kind of PCR fast reaction chip |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR100790881B1 (en) * | 2006-07-06 | 2008-01-02 | 삼성전자주식회사 | Micro-fluid reaction chip, and method for manufacturing the same |
WO2011138676A2 (en) * | 2010-05-04 | 2011-11-10 | King Abdullah University Of Science And Technology | Integrated microfluidic sensor system with magnetostrictive resonators |
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR100790881B1 (en) * | 2006-07-06 | 2008-01-02 | 삼성전자주식회사 | Micro-fluid reaction chip, and method for manufacturing the same |
WO2011138676A2 (en) * | 2010-05-04 | 2011-11-10 | King Abdullah University Of Science And Technology | Integrated microfluidic sensor system with magnetostrictive resonators |
Non-Patent Citations (1)
Title |
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A MULTIPORT METERING VALVE TECHNOLOGY FOR ON-CHIP VALVING(16th ICMS for CLS, Oct.28, 2012)* |
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