CN109355283B - Automatic nucleic acid extraction device suitable for space - Google Patents
Automatic nucleic acid extraction device suitable for space Download PDFInfo
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- CN109355283B CN109355283B CN201811422341.2A CN201811422341A CN109355283B CN 109355283 B CN109355283 B CN 109355283B CN 201811422341 A CN201811422341 A CN 201811422341A CN 109355283 B CN109355283 B CN 109355283B
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- injector
- nucleic acid
- stepping motor
- syringe
- rubber tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
Abstract
The invention discloses an automatic nucleic acid extraction device suitable for space, which comprises a translation stage, a stepping motor, a limit switch, an injector group, a silicone rubber tube, a pinch valve, an electromagnet and a control circuit. The injector group is five 1ml injectors fixedly connected together and is used for storing reagents required by nucleic acid extraction; the injector is fixed on a translation stage, and the translation stage is driven by a stepping motor and can push and pull a piston of the injector; the limit switch is positioned at the end of the piston stroke of the injector and is used for feeding back the position of the motor; all the syringes are connected together through a silicone rubber tube, and the pinch valve is used for controlling the on-off of the silicone rubber tube; two electromagnets are positioned at two sides of the main injector and are used for generating an oscillating magnetic field. The invention realizes the full-automatic extraction process from the sample to the purified nucleic acid in the liquid closed system, has no risk of aerosol pollution, can be used for automatically extracting the nucleic acid of the biological sample in the microgravity environment, and is convenient for integration with the subsequent nucleic acid processing module.
Description
Technical Field
The invention relates to a space biotechnology, in particular to a device which is suitable for a space microgravity environment and can realize automatic extraction of nucleic acid.
Background
Nucleic acids are an important object of modern molecular biological detection, the primary problem of which is the extraction of the desired nucleic acids from biological samples. The common nucleic acid extraction method in the laboratory comprises two types of manual extraction and automatic extraction, wherein the manual extraction needs to sequentially crack, adsorb, clean, elute and the like samples, the required steps are complicated, liquid is easy to volatilize in the air, and the samples are easy to pollute. The laboratory automatic nucleic acid extraction instrument is realized automatically in a machine through the step of manually extracting by a mechanical arm, and the used test tube is also open, so that the risk of sample pollution is also caused. Both of these methods are not suitable for use in a space microgravity environment.
Living plants, living animals and microorganisms are main objects of life science experiments in space stations in China, and at present, the research on molecular biology level of samples in the space life science experiments can be carried out only by returning the samples to the ground. The samples returned to the ground need to be fixed first, so that the samples are not influenced by the ground environment. Molecular biology research on the gene level is an important research means of modern living things and is indispensable for the research of space life science in China. Along with the construction of space stations in China, the space life science experiment needs to realize the extraction of nucleic acid in space. However, the nucleic acid extraction apparatus commonly used in the ground laboratory uses gravity to make the liquid at the bottom of the test tube, and operates the liquid under the condition of space microgravity, and the ground device cannot be directly used in space without a fixed gas-liquid interface. Meanwhile, complicated operation is performed in the space station, and the requirement on experimental operators is very high, so that the space station in China needs to research an automatic nucleic acid extraction technology suitable for space application.
Disclosure of Invention
The invention discloses an automatic nucleic acid extraction device suitable for space, which comprises an injector group, a fixed plate, a translation table, a stepping motor, a limit switch, a silicon rubber tube, a pinch valve, an electromagnet and a control circuit, and is characterized in that:
the stepping motors are of the same type and can ensure that the push-pull distance generated by each pulse is consistent;
the injector group is a 1ml slim tube injector which is fixedly connected together and made of polypropylene with good biocompatibility, and the injectors are fixedly connected together through the fixing plate and can be conveniently disassembled;
the fixing plate is a mounting plate for fixing the injector, a groove capable of being embedded into the injector tube is formed in the fixing plate, and the injector is fixed between the two clamping plates through screws;
the translation table is a miniature translation table, and the stroke can cover the movable range of the injector piston;
the pinch valve is a normally closed micro pinch valve, so that liquid interference among liquid exchanging steps can be reduced to the minimum;
the electromagnets are two micro electromagnets with the same model and a suction force of more than 2N, and can drive the magnetic beads in the injector to do reciprocating oscillation;
the injector group is connected through the silicon rubber tube and is arranged on the translation stage, and then the steps of sample splitting, cleaning, eluting and the like are sequentially realized by controlling the push-pull of the stepping motor and the on-off of the pinch valve.
The technical scheme for realizing the invention is as follows:
(1) The syringes filled with magnetic beads, pyrolysis liquid, cleaning liquid 1, cleaning liquid 2 and eluent are respectively and fixedly arranged on a fixed plate, are connected with a silicone rubber tube, and are packaged and stored by a clamping tube clamp;
(2) During the nucleic acid extraction operation, the sample is sucked into a syringe filled with magnetic beads, so as to finish sample addition; then mounting the syringe group after sample addition on a translation stage base;
(3) Starting an automatic nucleic acid extraction program, sequentially opening all pressure tube valves, pushing and pulling the injector to complete the operations of cracking, adsorbing, cleaning and eluting, and using an electromagnet to perform oscillating mixing or adsorbing on the liquid in the injector 3-1.
(4) The finally purified nucleic acid is positioned in the injector 3-2 and can be used for subsequent nucleic acid detection tests.
(5) After the whole purification test is finished, the syringe group is taken down, and the next nucleic acid extraction test is prepared.
The invention has the advantages that:
(1) In the nucleic acid extraction process, all liquid operations are completed in a sealed system, and the technology enables automatic extraction of nucleic acid to be performed in a microgravity environment;
(2) The vibration of the magnetic beads for automatically extracting the nucleic acid is realized by externally applying an alternating magnetic field, and the reciprocating vibration of the magnetic beads ensures that the mixing efficiency of the sample is higher;
(3) The closed system of the invention avoids the pollution of the nucleic acid sample, and simultaneously avoids the pollution of the operated sample to the environment, so that the nucleic acid operation can be completed in the same laboratory, and the probability of false positive is reduced.
(4) The device adopts the four-hole needle on the sample injector, reduces the liquid dead volume of the needle part, leads out the injector at the rear end of the silicon rubber pipeline respectively, and avoids the crosstalk problem of 4 liquids at the rear end.
(5) The injector used by the device has low cost, and is connected together through the external fixing plate, so that the injector is convenient to assemble, disassemble and replace.
Drawings
FIG. 1 is a schematic diagram showing the structure of an automatic nucleic acid extraction apparatus system suitable for use in space.
In the figure: 1. a stepping motor; 2. a translation stage; 3. a syringe set; 4. fixedly connecting a clamping plate; 5. a limit switch; 6. a silicone rubber tube; 7. a pinch valve; 8. an electromagnet.
Fig. 2 is a schematic view of a syringe assembly fixedly connected together.
In the figure: 3-1, a sample injector; 3-2 eluent syringe; 3-3 a cleaning liquid 2 syringe; 3-4 cleaning fluid 1 syringe; 3-5 lysate injector; 3-6 four-hole needle heads; and 4, fixing the plate.
Detailed Description
The device of the patent is verified in a laboratory, and the specific operation steps are as follows:
(1) Sucking the sample and the magnetic beads into the syringe 3-1 at the sterile operation table, sucking the lysate into the syringe 3-5, sucking the cleaning liquid 1 into the syringe 3-4, sucking the cleaning liquid 2 into the syringe 3-3, and sucking the eluent into the syringe 3-2; connecting the silicone rubber tubes 6, and clamping each silicone rubber tube by using a clamp; the syringe is mounted on the fixing plate 4, and the sample preparation is completed.
(2) After the pinch valve is powered on and opened, the connected injector group is mounted on the translation stage, and after the pinch valve is closed, the clamp used during sample adding is taken down, so that the mounting is completed; automatic nucleic acid extraction is initiated.
(3) Program control lysate injector 3-5 corresponding pressure pipe valve opens, the stepping motor promotes the injector 3-5, pulls the injector 3-1 at the same time, until the lysate in the injector 3-5 is completely injected into the injector 3-1, finish the injection of lysate; the electromagnet 8 is started to enter an oscillation mode, the oscillation time is determined according to the reagent requirement, and the cracking operation is completed.
(4) After cracking, the electromagnet 8 enters an adsorption mode, then the stepping motor pushes the injector 3-1, and simultaneously pulls the cracking liquid injector 3-5 until the piston in the injector 3-1 moves to the bottom, and closes the corresponding pressure pipe valve of the injector 3-5, so that the adsorption operation is completed.
(5) The program controls the valve of the injector 3-4 to open, the stepping motor pushes the injector 3-4 of the cleaning liquid 1, and pulls the injector 3-1 at the same time until the cleaning liquid 1 in the injector 3-4 is completely injected into the injector 3-1, and the suction of the cleaning liquid 1 is completed; the electromagnet 8 is started to enter an oscillation mode, the oscillation time is determined according to the reagent requirement, and the cleaning 1 operation is completed.
(6) After the cleaning 1 is finished, the electromagnet 8 enters an adsorption mode, then the stepping motor pushes the injector 3-1, and simultaneously pulls the cleaning liquid injector 3-4 until the piston in the injector 3-1 moves to the bottom, the cleaning liquid 1 is discharged, and the corresponding pinch valve of the injector 3-4 is closed.
(7) The program controls the valve of the pressure tube corresponding to the injector 3-3 to open, the stepping motor pushes the injector 3-3 of the cleaning liquid 2, and pulls the injector 3-1 at the same time until the cleaning liquid 2 in the injector 3-3 is completely injected into the injector 3-1, and the suction of the cleaning liquid 2 is completed; the electromagnet 8 is started to enter an oscillation mode, the oscillation time is determined according to the reagent requirement, and the cleaning operation of 2 is completed.
(8) After the cleaning 2 is finished, the electromagnet 8 enters an adsorption mode, then the stepping motor pushes the injector 3-1, and simultaneously pulls the cleaning liquid injector 3-3 until a piston in the injector 3-1 moves to the bottom, the cleaning liquid 2 is discharged, and the corresponding pinch valve of the injector 3-3 is closed.
(9) The program controls the valve of the corresponding plunger valve of the injector 3-2 to open, the stepping motor pushes the elution injector 3-2, and pulls the injector 3-1 at the same time until the eluent in the injector 3-2 is completely injected into the injector 3-1, thus finishing the inhalation of the eluent; the electromagnet 8 is started to enter an oscillation mode, the oscillation time is determined according to the reagent requirement, and the eluent inhalation operation is completed.
(10) After the elution is completed, the electromagnet 8 enters an adsorption mode, then the stepping motor pushes the injector 3-1, simultaneously pulls the eluent injector 3-2, and sucks the eluted purified nucleic acid into the injector 3-2. Closing the corresponding pressure tube valve of the injector 3-2 to finish nucleic acid extraction.
(11) The purified nucleic acid is amplified and verified by a real-time PCR instrument, and the device has the same nucleic acid extraction efficiency as a laboratory commercial nucleic acid extractor.
Claims (1)
1. The utility model provides an automatic extraction element of nucleic acid suitable for space, includes step motor (1), translation platform (2), syringe group (3), links firmly splint (4), limit switch (5), silicone rubber tube (6), pinch valve (7), electro-magnet (8) and control circuit, its characterized in that:
the stepping motor (1) is a miniature stepping motor of the same type, and can ensure that the push-pull distance generated by each pulse is consistent;
the syringe group (3) is a 1ml thin tube syringe made of polypropylene with good biocompatibility, which are fixedly connected together, the syringes can be conveniently disassembled and assembled by being fixedly connected together through the fixedly connected clamping plates (4), and the needle of the syringe is a four-hole needle;
the fixed connecting clamping plate (4) is a mounting plate for fixing the injector, a groove capable of being embedded into the injector tube is formed in the fixed connecting clamping plate, and the injector is fixed between the two clamping plates through screws;
the translation table (2) is a miniature translation table, and the stroke can cover the movable range of the piston of the injector group (3);
the pressure pipe valve (7) is a normally closed micro pressure pipe valve, so that the liquid interference among the liquid exchanging steps can be reduced to the minimum;
the electromagnet (8) is two micro electromagnets with the same model and a suction force larger than 2N, and can drive the magnetic beads in the injector (3-1) to do reciprocating oscillation;
the injector group (3) is connected through a silicon rubber tube (6) and is arranged on the translation table (2), and then the steps of sample splitting, cleaning and eluting are sequentially realized by controlling the push-pull of the stepping motor (1) and the on-off of the pinch valve (7).
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811422341.2A CN109355283B (en) | 2018-11-27 | 2018-11-27 | Automatic nucleic acid extraction device suitable for space |
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| CN201811422341.2A CN109355283B (en) | 2018-11-27 | 2018-11-27 | Automatic nucleic acid extraction device suitable for space |
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| CN109355283A CN109355283A (en) | 2019-02-19 |
| CN109355283B true CN109355283B (en) | 2023-05-05 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110628569A (en) * | 2019-10-16 | 2019-12-31 | 宁波美康盛德生物科技有限公司 | Full-automatic nucleic acid extraction system |
| CN111548926B (en) * | 2020-02-28 | 2021-06-22 | 青岛英赛特生物科技有限公司 | Totally-enclosed nucleic acid extraction and detection integrated kit |
| CN114570449B (en) * | 2022-04-26 | 2022-08-16 | 广州国家实验室 | Liquid transfer device and multi-path parallel liquid transfer device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232469A (en) * | 2014-09-11 | 2014-12-24 | 北京化工大学 | Magnetic bead-based sample treatment and nucleic acid automatic extraction system |
| CN104762193A (en) * | 2015-04-21 | 2015-07-08 | 北京化工大学 | Microfluidic device for automatic extraction of nucleic acid |
| CN108841818A (en) * | 2018-03-13 | 2018-11-20 | 融智生物科技（青岛）有限公司 | A kind of method for extracting nucleic acid and device |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| GB2416030B (en) * | 2004-01-28 | 2008-07-23 | Norchip As | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
| GB0710957D0 (en) * | 2007-06-07 | 2007-07-18 | Norchip As | A device for carrying out cell lysis and nucleic acid extraction |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232469A (en) * | 2014-09-11 | 2014-12-24 | 北京化工大学 | Magnetic bead-based sample treatment and nucleic acid automatic extraction system |
| CN104762193A (en) * | 2015-04-21 | 2015-07-08 | 北京化工大学 | Microfluidic device for automatic extraction of nucleic acid |
| CN108841818A (en) * | 2018-03-13 | 2018-11-20 | 融智生物科技（青岛）有限公司 | A kind of method for extracting nucleic acid and device |
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