WO2018053783A1 - 一种用于制备基因测序样本的系统及其应用 - Google Patents

一种用于制备基因测序样本的系统及其应用 Download PDF

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Publication number
WO2018053783A1
WO2018053783A1 PCT/CN2016/099816 CN2016099816W WO2018053783A1 WO 2018053783 A1 WO2018053783 A1 WO 2018053783A1 CN 2016099816 W CN2016099816 W CN 2016099816W WO 2018053783 A1 WO2018053783 A1 WO 2018053783A1
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WIPO (PCT)
Prior art keywords
module
porous plate
nucleic acid
heating
sample
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PCT/CN2016/099816
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English (en)
French (fr)
Inventor
李景
高建东
邹良英
刘慧�
李琳
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深圳华大智造科技有限公司
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Application filed by 深圳华大智造科技有限公司 filed Critical 深圳华大智造科技有限公司
Priority to CN201680081102.8A priority Critical patent/CN108603155A/zh
Priority to PCT/CN2016/099816 priority patent/WO2018053783A1/zh
Publication of WO2018053783A1 publication Critical patent/WO2018053783A1/zh

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00Apparatus for enzymology or microbiology
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00Apparatus for enzymology or microbiology
    • C12M1/36Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38Temperature-responsive control

Definitions

  • the present application relates to the field of gene sequencing, and more particularly to a system for preparing a gene sequencing sample and an application thereof.
  • DNA sequencing refers to the analysis of the base sequence of a specific DNA fragment, that is, the arrangement of adenine (A), thymine (T), cytosine (C) and guanine (G). the way.
  • RNA sequencing refers to the analysis of the base sequence of a particular RNA fragment, ie, the arrangement of adenine (A), guanine (G), cytosine (C), and uracil (U).
  • a nucleic acid molecule Before performing gene sequencing, it is necessary to extract a nucleic acid molecule from a source material containing a gene molecule, such as a plasma, a tissue sample, or a cell culture, and then amplify the nucleic acid molecule, usually by using a polymerase chain.
  • the reaction abbreviated PCR
  • the reaction is amplified, and then the nucleic acid molecule is purified, and finally the gene-sequenced library is obtained, which is used as a gene sequencing sample for sequencing by a sequencer.
  • the entire process from nucleic acid extraction, PCR amplification to purification to obtain a sequencing library is collectively referred to as gene sequencing sample preparation.
  • nucleic acid extraction requires multiple use of the centrifuge. After each centrifugation, the centrifuge tube is manually removed, the supernatant is removed or retained, and the reagent is added for centrifugation, and thus repeated.
  • the PCR amplification process also requires manual configuration of the reaction solution, setting the reaction procedure on the PCR instrument, and then manually adding the prepared DNA to the reaction solution, manually placing it on the PCR instrument for amplification, and after the amplification is completed, It was manually taken out, and a reagent was added to the amplification product for purification.
  • nucleic acid purification requires the use of other equipment and reagents.
  • One aspect of the present application discloses a system for preparing a genetically sequenced sample, comprising a sealed cavity surrounded by a housing 1, and a raw material placement module 2, a nucleic acid amplification module disposed within the sealed cavity. 3.
  • the heating and cooling module 4 the pipetting module, the magnetic bead purification module 6, the waste recycling module and the control module; the top end of the outer casing 1 is provided with an air inlet 11 and the air filtering unit 12 is installed in the air inlet 11 to The air in the sealed chamber is ensured to be clean;
  • the pipetting module includes an automatic pipetting gun 51 and a mechanical arm 52.
  • the automatic pipetting gun 51 is disposed on the robot arm 52.
  • the robot arm 52 controls the automatic pipetting gun 51 to move, and the automatic pipetting gun 51 For sucking and transferring a liquid sample;
  • the heating and cooling module 4 includes an electric heating unit, and a temperature-changing porous plate 41, a room temperature porous plate 42 disposed on the electric heating unit, and a row disposed under the temperature-changing porous plate 41 and the room temperature porous plate 42
  • the air fan 43 and the heat dissipating component 44, the electric heating component independently heats the variable temperature porous plate 41 and the normal temperature porous plate 42.
  • the variable temperature porous plate 41 can provide different according to the setting.
  • the nucleic acid amplification module 3 includes a heating sealing cap 31, a PCR perforated plate 32, a heat sink 33, and an exhaust fan 34.
  • the heating sealing cap 31 is subjected to a PCR reaction, and the PCR porous plate 32 is sealed and heated, and the PCR porous plate 32 is heated.
  • the radiator 33 and the exhaust fan 34 are used to control the reaction temperature;
  • the magnetic bead purification module 6 includes a purified porous plate 61, a holder 62, a magnetic frame 63 and a lifting mechanism 64, and the purified porous plate 61 is placed on the holder 62.
  • the magnetic frame 63 is independently disposed directly below the purified porous plate 61, and the lifting mechanism 64 controls the magnetic frame 63 to rise or fall independently;
  • the waste recycling module includes at least one waste collection box 71; and the control module and the nucleic acid amplification module 3, respectively
  • the heating and cooling module 4, the pipetting module signal connection, the independent control nucleic acid amplification module 3, the heating and cooling module 4 or the robot arm 52 operate.
  • the mechanical arm 52 is provided with three moving directions, namely an X-axis, a y-axis and a z-axis, and the robot arm 52 controls the automatic pipetting gun 51 to move in any of the X-axis, the y-axis and the z-axis.
  • the raw material placement module 2, the nucleic acid amplification module 3, the heating and cooling module 4, the magnetic bead purification module 6, and the waste collection module are sequentially arranged along the X axis of the robot arm 52.
  • the key of the present application is to integrate nucleic acid extraction, PCR amplification and magnetic bead purification into a system, and use a robotic arm to move an automatic pipette to perform reagent addition and sample transfer.
  • the process is carried out in an ultra-clean sealed chamber, avoiding manual errors or errors, and avoiding contamination of the genetically sequenced samples by environmental factors.
  • the reagent addition and sample transfer of the present application are all realized by the robot arm, therefore, the raw material placement module 2, the nucleic acid amplification module 3, the heating and cooling module 4, the magnetic bead purification module 6 and the waste recycling module are along the machinery.
  • the X-axis of the arm 52 is arranged in order to facilitate the movement sampling of the robot arm; as for the y-axis of the mechanical arm is horizontally perpendicular to the X-axis, and the X-axis is defined as the longitudinal direction, the y-axis is the realization of the mechanical arm in the width direction. Displacement, while the z-axis controls the automatic pipette to move up and down for aspiration sampling.
  • the raw material placement module of the present application is used for placing a liquid sample such as plasma or cell culture liquid, and a solid sample such as a tissue or a solid culture needs to be simply chopped or ground outside. Put the raw material placement module into the system of this application.
  • an ultraviolet sterilization lamp 9 is installed in the sealed cavity enclosed by the outer casing 1 for ultraviolet sterilization of the sealed cavity.
  • the ultraviolet sterilization lamp 9 is also installed in the system of the present application, as for the ultraviolet lamp.
  • the specific installation location can refer to equipment such as ultra-clean workbench.
  • a front viewable safety door 13 is mounted on the front side of the outer casing 1.
  • the two side faces of the outer casing 1 are provided with a see-through window 14.
  • the front, the side, the rear, and the like of the outer casing in the present application are defined in accordance with the surface of the operator in accordance with the test, that is, the front surface of the front operator is the front side.
  • both the see-through security door 13 and the see-through window 14 are made of a tinted transparent material.
  • the transparent security door 13 and the see-through window 14 are prepared by using a brown transparent material.
  • the ultraviolet rays can also be prevented from radiating outward. Understandably, others can reach The material of this function can be used in this application.
  • the pipetting module further comprises two tip placement modules 53, 54, for placing the tips.
  • the back side of the casing 1 is further provided with a USB interface 15, a serial port control interface 16, a power switch 17 and a network interface 18;
  • the power switch 17 controls the power supply of the entire system to be turned on or off;
  • the serial port control interface 16 is used to connect the control terminal, including a computer or other human-computer interaction interface;
  • the network interface 18 is used to connect the system to the network.
  • B-transfer interface 15 serial port control interface 16, power switch 17 and network interface 18, these interfaces can be easily connected to external devices, thus facilitating control and use.
  • the rear surface of the outer casing 1 is further provided with a heat dissipation opening 19.
  • the system of the present application further includes an external user terminal, by which the user terminal sends an instruction to the control module, and independently controls the raw material placement module 2, the nucleic acid amplification module 3, the heating and cooling module 4, and the pipetting module.
  • the magnetic bead purification module 6 and the waste recycling module are operated.
  • the user terminal may be a tester's own computer, and may be connected to the system of the present application through the USB transfer interface 15, the serial port control interface 16 or the network, and the corresponding software may be run on the computer;
  • the system has its own independent human-computer interaction interface to prevent the entire system from being interfered by other factors.
  • the system further comprises a scanner 10, which is mounted in a sealed cavity surrounded by the outer casing 1.
  • the reagent or various samples of the present application have a barcode or a two-dimensional code attached to the wall of the tube, and the corresponding sample and reagent information can be identified and recorded by scanning by the scanner 10, For sample tracking management, it can effectively improve operational efficiency.
  • Another aspect of the present application discloses the use of the system of the present application in nucleic acid extraction, nucleic acid purification, or library construction.
  • system of the present application is used for preparing a gene sequencing sample, which includes steps of nucleic acid extraction, PCR, magnetic bead purification, etc., of course, in some special cases, the system of the present application can also be used alone.
  • nucleic acid extraction nucleic acid purification or library construction.
  • the system for preparing a gene sequencing sample of the present application innovates and integrates various processes involved in preparing a gene sequencing sample, and realizes a one-stop automated sample preparation process, which is required to be performed after the sample is input.
  • library samples for sequencing can be exported.
  • the entire process is carried out in a clean sealed chamber, avoiding environmental pollution and cross-contamination between samples, and improving the quality and efficiency of preparing genetically sequenced samples.
  • FIG. 1 is a schematic diagram showing the overall appearance of a system for preparing a gene sequencing sample in the embodiment of the present application;
  • FIG. 2 is a system for preparing a gene sequencing sample in the embodiment of the present application, Schematic diagram of the internal structure
  • FIG. 3 is a schematic diagram of an internal structure of a system for preparing a genetic sequencing sample according to another embodiment of the present invention.
  • FIG. 4 is a schematic diagram of a back surface structure of a system for preparing a gene sequencing sample in the embodiment of the present application;
  • FIG. 5 is a system for preparing a gene sequencing sample in the embodiment of the present application, after removing the outer shell of the outer casing Schematic;
  • FIG. 6 is a schematic diagram of an internal structure of a system for preparing a gene sequencing sample in the embodiment of the present application, after removing the outer casing and the frame; [0043] FIG.
  • FIG. 7 is a schematic diagram showing the appearance of a heating and cooling module of a system for preparing a gene sequencing sample in an embodiment of the present application
  • FIG. 8 is a schematic diagram of an internal structure of a heating and cooling module for preparing a gene sequencing sample in the embodiment of the present application;
  • FIG. 9 is a schematic diagram showing the appearance of a magnetic bead purification module for preparing a gene sequencing sample in the embodiment of the present application.
  • FIG. 10 is a schematic diagram showing the decomposition structure of a magnetic bead purification module for preparing a gene sequencing sample in the embodiment of the present application;
  • FIG. 11 is a schematic diagram of a system for preparing a gene sequencing sample in the embodiment of the present application, the tip placement module of which is assembled with a suction head;
  • FIG. 12 is a system for preparing a gene sequencing sample in the embodiment of the present application, the tip placement module assembly suction Schematic diagram of the decomposition structure of the head;
  • FIG. 13 is a schematic view showing the structure of a nucleic acid amplification module for preparing a gene sequencing sample in the embodiment of the present application.
  • the system for preparing a gene sequencing sample of the present application the key is to integrate the various processes and equipment in the preparation process of the genetic sequencing sample into one system, the whole system is carried out in a relatively sealed cavity, and the whole is guaranteed The sealed chamber is clean.
  • the system for preparing a gene sequencing sample of the present application can realize automatic control of each module by control module control.
  • the system for preparing a gene sequencing sample of this example includes a sealed cavity surrounded by the outer casing 1, and a raw material placement module 2 disposed in the sealed cavity.
  • the top end of the outer casing 1 is provided with an air inlet 11 , and as shown in FIG. 5 , an air filtering unit 12 is installed in the air inlet 11 to ensure that the air in the sealed cavity is clean. Air enters through the air inlet 11 and is then filtered and purified by the air filter unit 12 to enter the interior of the system.
  • a front side of the outer casing 1 is mounted with a slidable see-through safety door 13, two sides of the outer casing 1 are provided with a see-through window 14; and the see-through security door 13 and the see-through window 14 are transparent in color.
  • Material preparation wherein, the see-through security door 13 can be pushed up and down, and the see-through security door 13 can be operated inside the system, for example, adding raw materials or taking out the prepared genetic sequencing sample, and closing the door after the system is running.
  • the brown transparent material is used to prepare the see-through security door 13 and the see-through window 14, which is convenient for the tester to observe the environment inside the system, and also prevents the ultraviolet ray of the ultraviolet sterilizing lamp 9 in the system from radiating outward.
  • the back of the casing 1 is further provided with a USB interface 15, a serial port control interface 16, a power switch 17, a network interface 18, and a heat dissipation port 19; the power switch 17 controls the power of the entire system to be turned on or off.
  • the USB transfer interface 15 and the serial port control interface 16 are used to connect the control terminal, including a computer or other human-computer interaction interface; the network interface 18 is used to connect the system to the network.
  • the raw material placement module 2 is disposed in the sealed cavity, and the raw material placement module 2 includes at least one raw material porous plate 21, and different samples can be placed in each of the raw material porous plates.
  • the pipetting module of this example includes an automatic pipetting gun 51, a robot arm 52, and two tip placement modules 53, 54.
  • Two nozzle placement modules 53, 54 are used for placing the suction head;
  • the robot arm 52 controls the automatic pipetting gun 51 to move arbitrarily in three directions of the X-axis, the y-axis and the z-axis;
  • the automatic pipetting gun 51 is used for suction and transfer Liquid sample.
  • the raw material placement module 2 the nucleic acid amplification module 3, the heating and cooling module 4, the magnetic bead purification module 6 and the waste recovery module are arranged in sequence along the X axis of the robot arm 52.
  • the raw material of this example is blood plasma.
  • the raw material placement module 2 is mainly used for placing liquid raw materials. For solid materials such as tissues, it needs to be crushed and homogenized before being placed in the system of the present application.
  • the two suction head placement modules 53, 54 are respectively placed with the tip porous plate 531, and a suction can be placed in each hole. Head 511.
  • the heating and cooling module 4 of the present example includes an electric heating unit, a temperature-changing porous plate 41 disposed on the electric heating unit, a room temperature porous plate 42, and a variable temperature porous plate 41. And the exhaust fan 43 and the heat dissipating component 44 below the normal temperature porous plate 42, the electric heating component independently heats the variable temperature porous plate 41 and the normal temperature porous plate 42, and the variable temperature porous plate 41 and the normal temperature porous plate 42 are used for placing different samples and reagents.
  • variable temperature porous plate 41 and the normal temperature porous plate 42 mainly rely on the electric heating unit to supply heat, and the exhaust fan 43 and the heat dissipating unit 44 are used for exhausting excess heat of the temperature-changing porous plate 41 and the normal temperature porous plate 42, so that the temperature-changing porous plate 41 and the room temperature are normal temperature.
  • the perforated plate 42 is capable of rapidly changing the temperature.
  • the room temperature porous plate 42 generally maintains a constant temperature.
  • the variable temperature porous plate 41 and the normal temperature porous plate 42 are both metal materials, and aluminum is specifically used in this example.
  • the nucleic acid amplification module 3 of the present example includes a heat sealing cover 31, a PCR porous plate 32, a heat sink 33, and an exhaust fan 34.
  • the PCR porous plate 32 is placed on the heat sink 33, and is heat-sealed.
  • the lid 31 is subjected to a P CR reaction, and the PCR porous plate 32 is tightly sealed and heated; the PCR porous plate 32 is used to place the reaction liquid, and the heat sealing cap 31 can supply heat to the PCR porous plate 32, control its temperature, and dissipate heat.
  • the fan 33 and the exhaust fan 34 are used to integrally control the reaction temperature in conjunction with the heat sealing cover 31.
  • the heat sink 33 is configured to remove heat from the PCR porous plate, the heat sink is provided with a plurality of heat dissipating fins 331, and one end of the heat dissipating fin is provided with an exhaust fan 34. To remove the heat from the fins.
  • the heat sealing cap 31 protrudes outwardly and covers the PCR porous plate 32.
  • the heat sealing cap can seal the pores to avoid evaporation of the sample or reagent in the pores, and can also be used for the pores.
  • the sample or reagent inside is accurately heated and cooled to provide a suitable temperature for the PCR reaction.
  • the magnetic bead purification module 6 of this example includes a purified porous plate 61, a support 62, a magnetic frame 63, and a lifting mechanism 64.
  • the purified porous plate 61 is placed on the support 62, and the magnetic force is applied.
  • the frame 63 is independently disposed directly below the purified porous plate 61, and the lifting mechanism 64 controls the magnetic frame 63 to rise or fall independently.
  • the magnetic frame 63 is used to provide a magnetic field, and the magnetic frame 63 is provided with a lifting structure 64, which can drive the magnetic frame 63 to move up and down rapidly.
  • the holder 62 is mainly used to support the purified porous plate 61, and the purified porous plate is filled with a magnetic bead solution.
  • the magnetic frame 63 and the bracket 62 are not integrated, but are separated.
  • the lifting structure 64 drives the magnetic frame 63 to move up and down
  • the bracket 62 directly supports the purified porous plate 61 and remains stationary.
  • the magnetic frame 63 is lowered, and the reaction liquid in the purified porous plate 61 is sucked/released multiple times through the suction head to achieve the purpose of mixing; when it is necessary to retain the magnetic beads, the liquid ⁇ is removed, and the magnetic frame 63 rises.
  • the magnetic beads are adsorbed to the bottom of the purified porous plate 61, and the liquid can be sucked away by the suction head to retain the magnetic beads, thereby cleaning and purifying the surface of the magnetic beads.
  • the waste recycling module of this example is a waste recycling box 71.
  • the control module is respectively connected to the nucleic acid amplification module 3, the heating and cooling module 4, the pipetting module signal, and the independent control nucleic acid amplification module 3, the heating and cooling module 4 or the mechanical arm 52.
  • the ultraviolet sterilization lamp 9 is installed in a sealed cavity enclosed by the outer casing 1 for ultraviolet sterilization of the sealed cavity.
  • the scanner 10 is mounted in a sealed cavity surrounded by the outer casing 1.
  • the reagent and various samples are affixed with a barcode or a two-dimensional code on the container, and can be identified and recorded by scanning by the scanner 10. Corresponding sample and reagent information for sample tracking management.
  • a conventional power supply module and a control module are provided. These can be referred to a conventional power supply and control module, which is not specifically limited herein, and is not specifically shown in the drawings.
  • the system for preparing a gene sequencing sample of the present example can be automatically controlled by a control module, and any operation program can be set as needed, and the use range is wide, and multiple different samples can be operated in the same manner.
  • the system for preparing a gene sequencing sample of this example one of which is specifically used as follows:
  • a raw material perforated plate is placed on the raw material placement module, and different plasma samples are placed in the pores of the porous plate of the raw material; the PCR porous plate may be preloaded with various reagents, or may not be loaded. Reagent; variable temperature porous plate and normal temperature porous plate are pre-packed with various reagent solutions; the porous plate of the tip is placed with a plurality of clean tips; the purified porous plate is pre-loaded with various required reagent solutions;
  • the robot arm moves the automatic pipetting gun to the top of the porous plate of the tip, the automatic pipetting gun moves downward and the required suction head is loaded and unloaded by pressure, and the pressure of the suction nozzle ensures the success of the loading and unloading nozzle and its sealing property. And then lift the tip up;
  • the robot arm moves the tip to the normal temperature porous plate or the purified porous plate, and the reagent is sucked into the suction head, and then the mechanical arm moves the suction head to the variable temperature porous plate to release the liquid in the suction head to The variable temperature is in the porous plate; the mechanical arm then moves the tip to the top of the porous plate of the tip, and releases the tip into the hole for subsequent recycling. If the tip is not needed later, the tip can be directly released. Discarded into the waste recycling box;
  • the mechanical arm moves the tip to the variable temperature porous plate, and the liquid in the tip is released into the variable temperature porous plate.
  • the system can repeat these steps to mix the plasma sample and various reagents, in the process, heating and cooling
  • the module can continuously heat or cool the variable temperature porous plate to provide a suitable temperature for the chemical reaction; the mechanical arm then moves the tip to the top of the porous plate of the tip, and releases the tip into the hole for subsequent recycling. If these tips are not needed afterwards, the tip release can be directly discarded into the waste recycling box;
  • the automatic pipette is loaded with a new tip, the liquid in the variable temperature porous plate is moved to the purified multi-well plate, the magnetic frame is raised, the magnetic beads are adsorbed on the bottom of the purified multi-well plate, and the automatic pipette sucks the waste liquid.
  • the robot arm then moves the tip over the perforated plate of the tip and releases the tip into the hole for subsequent recycling. If these tips are not needed later, the tip can be released directly to the waste recycling.
  • the magnetic frame is lowered, the cleaning liquid can be added to the hole with the magnetic beads by the suction head, and then the magnetic bead solution is repeatedly sucked/released, After cleaning the magnetic beads, the magnetic frame rises, the waste liquid is sucked away, and then the magnetic frame is lowered to re-add new reagents. Repeating this step can play the role of repeatedly cleaning the magnetic beads; finally, the magnetic frame rises and the magnetic beads are adsorbed.
  • the automatic pipette sucks away the waste liquid; leaving the magnetic beads;
  • the automatic pipette is moved to the purified porous plate to absorb the obtained DNA solution, and the extracted DNA solution is transferred to a PCR porous plate, and then moved to a normal temperature porous plate to absorb the enzyme reaction solution, and the automatic pipetting gun is moved to PCR multi-well plate, release the enzyme reaction solution in the tip, and then the DNA solution is reacted in the PCR multi-well plate; the robot arm moves the tip to the top of the porous plate of the tip, and releases the tip into the hole for subsequent circulation. Use, if you do not need to use these tips later, you can directly release the tip to the waste recycling box;
  • the DNA solution for completing the enzymatic reaction needs to be subjected to a purification operation: a magnetic bead solution is mixed into the DNA molecule solution, and after sufficient reaction, the DNA molecules are adsorbed on the magnetic beads, and then the automatic pipetting gun is exchanged for a new tip. Transferring the magnetic bead solution with the adsorbed DNA molecules to the purified multi-well plate; the mechanical arm then moves the tip to the top of the porous plate of the tip, and releases the tip into the hole for subsequent recycling, if it is not needed later Using these tips, the tip release can be directly discarded into the waste recycling box; the steps of purifying the magnetic beads can be referred to steps 9, 10;
  • DNA amplification After the DNA amplification is completed, it can be manually taken out, quantitatively processed, and then returned to the system for subsequent treatment, and various reagents are added to carry out various reactions to form a DNA nanosphere solution of a desired concentration. ;
  • the system of the present example can complete the database operation of DNA extraction, enzymatic reaction and purification by one-stop automation, and finally obtain the gene sequencing sample, and the specific example is the DNA nanosphere library.
  • the system of this example can be used with the sequencer developed by Huada Gene to provide genetic sequencing samples.
  • the raw material perforated plate of this example can hold 16 test samples, and the automatic pipetting gun can carry 8 suction heads at the same time. Of course, it can also be designed as needed according to needs.

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Abstract

本申请公开了一种用于制备基因测序样本的系统及其应用。本申请的系统,包括外壳围成的密封腔体,及设在密封腔体内的原材料放置模块、核酸扩增模块、加热制冷模块、移液模块、磁珠纯化模块、废料回收模块和控制模块;外壳顶端设进风口,安装空气过滤单元;移液模块包括自动移液枪和控制自动移液枪移动的机械臂;加热制冷模块包括变温多孔板、常温多孔板;核酸扩增模块包括加热密封盖、PCR多孔板、散热器和排风扇;磁珠纯化模块包括纯化多孔板、支架、磁力架和升降机构,控制模块独立控制各组件运行。本申请的系统,实现了一站式自动化样本制备,输入样本后可输出用于测序的文库样本,避免环境污染和样本交叉污染,提高了测序样本质量和效率。

Description

说明书 发明名称:一种用于制备基因测序样本的系统及其应用 技术领域
[0001] 本申请涉及基因测序领域, 特别是涉及一种用于制备基因测序样本的系统及其 应用。
[0002]
[0003] 背景技术
[0004] 基因测序, 或称核酸测序, 是研究核酸的重要方法之一, 包括 DNA测序和 RN A测序。 DNA测序 (DNA sequencing, 或译 DNA定序) 是指分析特定 DNA片段 的碱基序列, 也就是腺嘌呤 (A) 、 胸腺嘧啶 (T) 、 胞嘧啶 (C) 与鸟嘌呤 (G ) 的排列方式。 同样的, RNA测序是指分析特定 RNA片段的碱基序列, 也就是 腺嘌呤 (A) 、 鸟嘌呤 (G) 、 胞嘧啶 (C) 和尿嘧啶 (U) 的排列方式。
[0005] 在进行基因测序前, 需要先从含基因分子的源物质, 例如血浆、 组织样本或细 胞培养物等, 中提取核酸分子, 然后对核酸分子进行扩增, 通常是利用聚合酶 链式反应 (缩写 PCR) 进行扩增, 接着再对核酸分子进行纯化, 最后才能得到基 因测序的文库, 该测序文库作为基因测序样本, 用于测序仪进行测序。 从核酸 提取、 PCR扩增到纯化获得测序文库整个过程统称为基因测序样本制备。
[0006] 在基因测序样本的制备过程中, 核酸提取、 PCR扩增、 核酸纯化等是分别在不 同的仪器上, 由人工配合操作进行的。 例如核酸提取, 需要多次用到离心机, 每次离心完成后, 由人工取出离心管, 去除或保留上清液, 再加入试剂进行离 心, 如此反复。 而 PCR扩增过程也是需要人工配置好反应液, 在 PCR仪上设定反 应程序, 然后将制备的 DNA人工加入到反应液中, 人工放置于 PCR仪上进行扩 增, 扩增完成后, 再人工将其取出, 向扩增产物中加入试剂进行纯化。 同样的 , 核酸纯化又需要用到其它的仪器设备和试剂。
[0007] 可见, 制备基因测序样本的处理流程复杂、 耗吋; 而且由于试验过程对各种试 剂溶液的用量、 放置程序、 操作环境等有严格的要求, 而目前的处理方式几乎 都是人工操作完成, 难免出现错误或误差; 更重要的是, 在试剂的添加或者样 本的转移过程中, 人工操作容易造成交叉污染, 影响基因测序结果。 此外, 现 有的基因测序样本的处理流程中, 各试验阶段的设备都是单独完成特定试验环 节的, 例如离心、 PCR扩增等, 整个过程中需要人工操作进行转移, 程序复杂、 工作量大、 容易出错, 而且多台设备需要占用较大空间。
[0008]
[0009] 发明内容
[0010] 本申请的目的是提供一种用于制备基因测序样本的系统及其应用。
[0011] 为了实现上述目的, 本申请采用了以下技术方案:
[0012] 本申请的一方面公幵了一种用于制备基因测序样本的系统, 包括一个由外壳 1 围成的密封腔体, 以及设置在密封腔体内的原材料放置模块 2、 核酸扩增模块 3 、 加热制冷模块 4、 移液模块、 磁珠纯化模块 6、 废料回收模块和控制模块; 外 壳 1的顶端幵设有进风口 11, 并且, 于进风口 11内安装有空气过滤单元 12, 以保 障密封腔体内的空气干净; 移液模块包括自动移液枪 51和机械臂 52, 自动移液 枪 51设于机械臂 52上, 机械臂 52控制自动移液枪 51移动, 自动移液枪 51用于吸 取和转移液体样本; 加热制冷模块 4包括电加热组件, 和设置于电加热组件上的 变温多孔板 41、 常温多孔板 42, 以及设置于变温多孔板 41和常温多孔板 42下方 的排气扇 43和散热组件 44, 电加热组件独立的给变温多孔板 41、 常温多孔板 42 加热, 通常, 变温多孔板 41可以根据设定提供不同的温度, 而常温多孔板 42则 保持常温, 变温多孔板 41和常温多孔板 42用于放置不同的样本和试剂, 排气扇 4 3和散热组件 44用于控制变温多孔板 41和常温多孔板 42的温度; 核酸扩增模块 3 包括加热密封盖 31、 PCR多孔板 32、 散热器 33和排风扇 34, 加热密封盖 31在进行 PCR反应吋将 PCR多孔板 32密封, 并进行加热, PCR多孔板 32用于放置反应液, 散热器 33和排风扇 34用于控制反应温度; 磁珠纯化模块 6包括纯化多孔板 61、 支 架 62、 磁力架 63和升降机构 64, 纯化多孔板 61放置于支架 62之上, 磁力架 63独 立的设置于纯化多孔板 61的正下方, 并且, 升降机构 64控制磁力架 63独立上升 或下降; 废料回收模块包括至少一个废料回收盒 71 ; 控制模块分别与核酸扩增 模块 3、 加热制冷模块 4、 移液模块信号连接, 独立的控制核酸扩增模块 3、 加热 制冷模块 4或机械臂 52运行。 [0013] 优选的, 机械臂 52设置了三个移动方向, 即 X轴、 y轴和 z轴, 机械臂 52控制自 动移液枪 51在 X轴、 y轴和 z轴三个方向上任意移动; 与此同吋, 原材料放置模块 2 、 核酸扩增模块 3、 加热制冷模块 4、 磁珠纯化模块 6和废料回收模块, 沿机械臂 52的 X轴依序布置。
[0014] 需要说明的是, 本申请的关键在于将核酸提取、 PCR扩增和磁珠纯化等有机的 整合到一个系统中, 利用机械臂移动自动移液枪, 进行试剂添加和样本转移, 整个过程在一个超净密封腔体内进行, 避免了人工操作的错误或误差, 同吋也 避免了环境因素对基因测序样本的污染。 可以理解, 本申请的试剂添加和样本 转移都是由机械臂实现的, 因此, 原材料放置模块 2、 核酸扩增模块 3、 加热制 冷模块 4、 磁珠纯化模块 6和废料回收模块, 是沿机械臂 52的 X轴依序布置, 以方 便机械臂的移动取样; 至于机械臂的 y轴是水平垂直于 X轴的, X轴定义为长度方 向, 则 y轴就是实现机械臂在宽度方向上的位移, 而 z轴则是控制自动移液枪上下 移动, 以进行吸液取样。
[0015] 还需要说明的是, 本申请的原材料放置模块用于放置血浆或细胞培养液等液态 样本, 至于组织或固体培养物这样的固态样本, 需要在外面进行简单的切碎或 研磨后才能放入本申请系统的原材料放置模块。
[0016] 优选的, 本申请的系统中, 外壳 1围成的密封腔体内还安装有紫外灭菌灯 9, 用 于对密封腔体进行紫外灭菌消毒。
[0017] 可以理解, 除了采用空气过滤单元保持密封腔体内空气干净以外, 紫外线灭菌 消毒也是实验室中常用的, 因此, 在本申请的系统中同样安装有紫外灭菌灯 9, 至于紫外灯的具体安装位置可以参考超净工作台等设备。
[0018] 优选的, 外壳 1的前面安装有一个可幵合的透视安全门 13。
[0019] 优选的, 外壳 1的两个侧面幵设有透视窗口 14。
[0020] 需要说明的是, 本申请中外壳的前面、 侧面、 后面等是按照进行试验吋, 面对 操作人员的面而定义的, 也就是说, 正向操作人员的面即前面。
[0021] 优选的, 透视安全门 13和透视窗口 14都采用茶色透明材料制备。
[0022] 需要说明的是, 采用茶色透明材料制备透视安全门 13和透视窗口 14, 一方面是 为了方便观察, 同吋, 也可以避免紫外线向外辐射。 可以理解, 其它能够达到 该功能的材料都可以用于本申请。
[0023] 优选的, 移液模块还包括两个吸头放置模块 53、 54, 用于放置吸头。
[0024] 优选的, 外壳 1的背面还设置有 USB转接口 15、 串口控制接口 16、 电源幵关 17 和网络接口 18; 电源幵关 17控制整个系统的电源幵启或关闭; USB转接口 15和串 口控制接口 16用于连接控制终端, 包括电脑或其它人机交互界面; 网络接口 18 用于将系统与网络连接。
[0025] 需要说明的是, 为了方便操作使用, 本申请的系统, 在外壳 1的背面设置有 US
B转接口 15、 串口控制接口 16、 电源幵关 17和网络接口 18, 这些接口可以方便的 连接外部设备, 从而方便控制和使用。
[0026] 优选的, 外壳 1的背面还设置有散热口 19。
[0027] 优选的, 本申请的系统还包括外接的用户终端, 通过所述用户终端给控制模块 发送指令, 独立的控制原材料放置模块 2、 核酸扩增模块 3、 加热制冷模块 4、 移 液模块、 磁珠纯化模块 6和废料回收模块运行。
[0028] 需要说明的是, 用户终端可以是试验人员自己的电脑, 通过 USB转接口 15、 串 口控制接口 16或网络与本申请的系统连接, 在电脑上运行相应的软件即可; 也 可以是系统自带的独立的其它人机交互界面, 避免整个系统运行受到其它因素 的干扰。
[0029] 优选的, 系统还包括扫码器 10, 扫码器 10安装于由外壳 1围成的密封腔体内。
[0030] 需要说明的是, 本申请的试剂或各种样本, 在其管壁上都贴有条形码或者二维 码, 通过扫码器 10扫描就能够识别、 记录对应的样本和试剂信息, 用于样本跟 踪管理, 能够有效提高操作效率。
[0031] 本申请的另一面公幵了本申请的系统在核酸提取、 核酸纯化或文库构建中的应 用。
[0032] 可以理解, 本申请的系统是用于制备基因测序样本的, 其中包含了核酸提取、 PCR、 磁珠纯化等步骤, 当然, 在一些特殊情况下, 也可以将本申请的系统单独 用于核酸提取、 核酸纯化或文库构建等。
[0033]
[0034] 由于采用以上技术方案, 本申请的有益效果在于: [0035] 本申请的用于制备基因测序样本的系统, 对制备基因测序样本过程中涉及的各 个流程进行创新整合, 实现了一站式的自动化样本制备过程, 在输入样本后, 除了中途需要进行人工定量外, 可以输出用于测序的文库样本。 并且, 整个过 程在干净的密封腔体内进行, 避免了环境污染和样本之间的交叉污染, 提高了 制备基因测序样本的质量和效率。
[0036]
[0037] 附图说明
[0038] 图 1是本申请实施例中用于制备基因测序样本的系统的整体外观结构示意图; [0039] 图 2是本申请实施例中用于制备基因测序样本的系统, 打幵透视安全门看到的 内部结构示意图;
[0040] 图 3是本申请实施例中用于制备基因测序样本的系统, 打幵透视安全门另一视 角看到的内部结构示意图;
[0041] 图 4是本申请实施例中用于制备基因测序样本的系统, 其背面结构示意图; [0042] 图 5是本申请实施例中用于制备基因测序样本的系统, 去除外壳后的内部结构 示意图;
[0043] 图 6是本申请实施例中用于制备基因测序样本的系统, 去除外壳和框架后的内 部结构示意图;
[0044] 图 7是本申请实施例中用于制备基因测序样本的系统, 其加热制冷模块的外观 结构示意图;
[0045] 图 8是本申请实施例中用于制备基因测序样本的系统, 其加热制冷模块的内部 结构示意图;
[0046] 图 9是本申请实施例中用于制备基因测序样本的系统, 其磁珠纯化模块的外观 结构示意图;
[0047] 图 10是本申请实施例中用于制备基因测序样本的系统, 其磁珠纯化模块的分解 结构示意图;
[0048] 图 11是本申请实施例中用于制备基因测序样本的系统, 其吸头放置模块装配吸 头的结构示意图;
[0049] 图 12是本申请实施例中用于制备基因测序样本的系统, 其吸头放置模块装配吸 头的分解结构示意图;
[0050] 图 13是本申请实施例中用于制备基因测序样本的系统, 其核酸扩增模块的结构 示意图。
[0051]
[0052] 具体实施方式
[0053] 本申请的用于制备基因测序样本的系统, 其关键在于将基因测序样本制备过程 中的各个流程和设备整合到一个系统中, 整个系统在一个相对密封的腔体内进 行, 并且保障整个密封腔体干净。 本申请的用于制备基因测序样本的系统, 通 过控制模块控制, 可以实现各个模块的自动化控制。
[0054]
[0055] 下面通过具体实施例和附图对本申请作进一步详细说明。 以下实施例仅对本申 请进行进一步说明, 不应理解为对本申请的限制。
[0056]
[0057] 实施例
[0058] 本例的用于制备基因测序样本的系统, 如图 1至图 6所示, 包括一个由外壳 1围 成的密封腔体, 以及设置在密封腔体内的原材料放置模块 2、 核酸扩增模块 3、 加热制冷模块 4、 移液模块、 磁珠纯化模块 6、 废料回收模块、 紫外灭菌灯 9、 扫 码器 10和控制模块。 如图 1所示, 外壳 1的顶端幵设有进风口 11, 并且, 如图 5所 示, 于进风口 11内安装有空气过滤单元 12, 以保障密封腔体内的空气干净, 夕卜 界的空气通过进风口 11进入, 然后经过空气过滤单元 12过滤净化后进入系统内 部。 如图 2和图 3所示, 外壳 1的前面安装有一个可幵合的透视安全门 13, 外壳 1 的两个侧面幵设有透视窗口 14; 并且, 透视安全门 13和透视窗口 14都采用茶色 透明材料制备。 其中, 透视安全门 13可以上下推拉, 透视安全门 13打幵吋可对 系统内部进行操作, 例如添加原材料或者将制备好的基因测序样本取出, 系统 运行吋则关闭此门。 茶色透明材料制备透视安全门 13和透视窗口 14, 既可以方 便试验人员观察系统内环境, 同吋还能够阻止系统内紫外灭菌灯 9的紫外线向外 辐射。 如图 4所示, 外壳 1的背面还设置有 USB转接口 15、 串口控制接口 16、 电源 幵关 17、 网络接口 18和散热口 19; 电源幵关 17控制整个系统的电源幵启或关闭 ; USB转接口 15和串口控制接口 16用于连接控制终端, 包括电脑或其它人机交互 界面; 网络接口 18用于将系统与网络连接。 原材料放置模块 2设置于密封腔体内 , 并且, 原材料放置模块 2包括至少一个原料多孔板 21, 每个原料多孔板内可以 放置不同的样本。
[0059] 本例的移液模块如图 5和图 6所示, 包括自动移液枪 51、 机械臂 52和两个吸头放 置模块 53、 54。 两个吸头放置模块 53、 54用于放置吸头; 机械臂 52控制自动移 液枪 51在 X轴、 y轴和 z轴三个方向上任意移动; 自动移液枪 51用于吸取和转移液 体样本。 如图 6所示, 原材料放置模块 2、 核酸扩增模块 3、 加热制冷模块 4、 磁 珠纯化模块 6和废料回收模块, 沿机械臂 52的 X轴依序布置。 本例的原材料为血 浆, 本例中原材料放置模块 2主要是用于放置液态原材料, 至于组织等固态的原 材料, 则需要将其打碎匀浆后才能放入本申请的系统中。 本例的两个吸头放置 模块 53、 54, 如图 11和图 12所示, 两个吸头放置模块 53、 54上都分别放置了吸 头多孔板 531, 每个孔内可以放置一个吸头 511。
[0060] 本例的加热制冷模块 4, 如图 7和图 8所示, 包括电加热组件, 和设置于电加热 组件上的变温多孔板 41、 常温多孔板 42, 以及设置于变温多孔板 41和常温多孔 板 42下方的排气扇 43和散热组件 44, 电加热组件独立的给变温多孔板 41和常温 多孔板 42加热, 变温多孔板 41和常温多孔板 42用于放置不同的样本和试剂, 变 温多孔板 41和常温多孔板 42主要依靠电加热组件提供热量, 排气扇 43和散热组 件 44用于排走变温多孔板 41和常温多孔板 42多余的热量, 使得变温多孔板 41和 常温多孔板 42能够迅速改变温度。 其中, 常温多孔板 42—般保持一个不变的温 度, 为了能够快速控制温度, 变温多孔板 41和常温多孔板 42均为金属材料, 本 例具体的采用铝。
[0061] 本例的核酸扩增模块 3, 如图 13所示, 包括加热密封盖 31、 PCR多孔板 32、 散 热器 33和排风扇 34, PCR多孔板 32放置于散热器 33之上, 加热密封盖 31在进行 P CR反应吋将 PCR多孔板 32盖紧密封, 并进行加热; PCR多孔板 32用于放置反应 液, 加热密封盖 31能够为 PCR多孔板 32提供热量, 控制它的温度, 散热器 33和排 风扇 34用于配合加热密封盖 31整体控制反应温度。 其中, 散热器 33用于排走 PCR 多孔板的热量, 散热器设有多个散热翅片 331, 散热翅片一端设有排风扇 34, 用 于排走翅片内的热气。 在进行 PCR扩增吋, 加热密封盖 31向外伸出并盖合于 PCR 多孔板 32上, 加热密封盖可以对孔进行密封, 避免孔内样本或试剂的蒸发, 同 吋, 还可以对孔内的样本或试剂进行精准加温制冷, 为 PCR反应提供合适的温度
[0062] 本例的磁珠纯化模块 6, 如图 9和图 10所示, 包括纯化多孔板 61、 支架 62、 磁力 架 63和升降机构 64, 纯化多孔板 61放置于支架 62之上, 磁力架 63独立的设置于 纯化多孔板 61的正下方, 并且, 升降机构 64控制磁力架 63独立上升或下降。 其 中, 磁力架 63用于提供磁场, 磁力架 63设有升降结构 64, 可以驱动磁力架 63上 下快速移动。 支架 62主要是用于支撑纯化多孔板 61, 纯化多孔板内装有磁珠溶 液。 需要注意的是, 磁力架 63与支架 62并不是一体的, 而是分离的, 这样, 当 升降结构 64驱动磁力架 63上下移动吋, 支架 62—直支撑着纯化多孔板 61并且保 持不动, 当需要混匀磁珠吋, 磁力架 63下降, 通过吸头多次吸取 /释放纯化多孔 板 61内的反应液, 达到混匀的目的; 当需要保留磁珠, 去除液体吋, 磁力架 63 上升, 磁珠会吸附到纯化多孔板 61底部, 可以使用吸头吸走液体, 保留磁珠, 起到清洗纯化磁珠表面的效果。
[0063] 本例的废料回收模块为一个废料回收盒 71。 控制模块分别与核酸扩增模块 3、 加热制冷模块 4、 移液模块信号连接, 独立的控制核酸扩增模块 3、 加热制冷模 块 4或机械臂 52运行。 紫外灭菌灯 9安装于外壳 1围成的密封腔体内, 用于对密封 腔体进行紫外灭菌消毒。 扫码器 10安装于由外壳 1围成的密封腔体内, 本例中, 试剂和各种样本, 在其容器上都贴有条形码或二维码, 通过扫码器 10扫描就能 够识别、 记录对应的样本和试剂信息, 用于样本跟踪管理。 本例的系统内设置 有常规的电源模块和控制模块, 这些可以参考常规的电源和控制模块, 在此不 做具体限定, 图中也未具体示出。
[0064] 本例的制备基因测序样本的系统可由控制模块自动化控制, 可根据需要设定任 意操作程序, 使用范围广, 可实现同吋操作多个不同的样本。 本例的制备基因 测序样本的系统, 其中一种具体使用方法如下:
[0065] 1.系统处于原始状态, 使用前可以幵启紫外灭菌灯和空气过滤单元对系统的密 封腔体进行灭菌消毒; [0066] 2.打幵透视安全门, 将各种多孔板、 试剂和样本放入对应的模块中, 放入这些 试剂和样本以前先在扫码器处识别编码, 将试剂和样本的信息录入系统, 便于 系统自动化识别;
[0067] 3.在本实施例中, 原材料放置模块上放置有原材料多孔板, 原材料多孔板的孔 内放有不同的血浆样本; PCR多孔板可以预先装入各种试剂, 也可以不装入试剂 ; 变温多孔板和常温多孔板内预装各种所需的试剂溶液; 吸头多孔板放置有多 个干净的吸头; 纯化多孔板预先装入各种所需试剂溶液;
[0068] 4.关闭透视安全门, 并向系统输入执行程序, 启动系统运作;
[0069] 5.机械臂将自动移液枪移动至吸头多孔板上方, 自动移液枪向下移动并通过压 力装卸所需的吸头, 通过压力按压保证装卸吸头成功及它的密封性, 然后再向 上提起吸头;
[0070] 6.机械臂将吸头移至常温多孔板或纯化多孔板上方, 并将试剂吸取至吸头内, 然后机械臂再将吸头移至变温多孔板, 将吸头内液体释放到变温多孔板内; 机 械臂再将吸头移至吸头多孔板上方, 并将吸头释放到孔内以备后续循环利用, 若后续不需要再使用这些吸头, 则可以直接将吸头释放丢弃至废料回收盒内;
[0071] 7.自动移液枪装载新的吸头, 然后机械臂将吸头移至原材料多孔板并吸取血浆 样本;
[0072] 8.机械臂将吸头移至变温多孔板, 将吸头内液体释放到变温多孔板内, 系统可 以反复这些步骤, 将血浆样本和各种试剂混合, 在这个过程中, 加热制冷模块 可以不断对变温多孔板进行加热或者制冷的操作, 为化学反应提供合适的温度 ; 机械臂再将吸头移至吸头多孔板上方, 并将吸头释放到孔内以备后续循环利 用, 若后续不需要再使用这些吸头, 则可以直接将吸头释放丢弃至废料回收盒 内;
[0073] 9.自动移液枪装载新的吸头, 将变温多孔板内的液体移至纯化多孔板, 磁力架 上升, 将磁珠吸附在纯化多孔板底部, 自动移液枪吸走废液; 机械臂再将吸头 移至吸头多孔板上方, 并将吸头释放到孔内以备后续循环利用, 若后续不需要 再使用这些吸头, 则可以直接将吸头释放丢弃至废料回收盒内; 磁力架下降, 可以利用吸头向有磁珠的孔内加入清洗液, 然后反复吸取 /释放磁珠溶液, 起到 清洗磁珠的作用, 然后磁力架上升, 吸走废液, 接着磁力架下降重新加入新的 试剂, 反复本步骤, 则可以起到反复清洗磁珠的作用; 最后磁力架上升, 将磁 珠吸附在纯化多孔板底部, 自动移液枪吸走废液; 剩下磁珠;
[0074] 10.磁力架下降, 自动移液枪移至纯化多孔板吸取试剂, 将试剂释放到有磁珠的 孔位, 反复吸取 /释放磁珠溶液, 将磁珠上吸附的 DNA洗脱下来; 然后磁力架上 升, 将磁珠吸附在孔的底部, 这样就得到 DNA分子的溶液, 这些操作可以最终 从血浆中提取出 DNA分子; 机械臂再将吸头移至吸头多孔板上方, 并将吸头释 放到孔内以备后续循环利用, 若后续不需要再使用这些吸头, 则可以直接将吸 头释放丢弃至废料回收盒内;
[0075] 11.自动移液枪移至纯化多孔板吸取得到的 DNA溶液, 将提取后的 DNA溶液移 至 PCR多孔板内, 然后移至常温多孔板吸取酶反应液, 自动移液枪移至 PCR多孔 板, 释放吸头内的酶反应液, 然后 DNA溶液在 PCR多孔板内进行反应; 机械臂 再将吸头移至吸头多孔板上方, 并将吸头释放到孔内以备后续循环利用, 若后 续不需要再使用这些吸头, 则可以直接将吸头释放丢弃至废料回收盒内;
[0076] 12.完成酶反应的 DNA溶液, 需要进行纯化操作: 向 DNA分子溶液混入磁珠溶 液, 充分反应后, DNA分子吸附于磁珠上, 然后自动移液枪换取新的吸头之后 , 将该吸附有 DNA分子的磁珠溶液转移至纯化多孔板内; 机械臂再将吸头移至 吸头多孔板上方, 并将吸头释放到孔内以备后续循环利用, 若后续不需要再使 用这些吸头, 则可以直接将吸头释放丢弃至废料回收盒内; 纯化磁珠的步骤可 参考步骤 9、 10;
[0077] 13.将纯净的 DNA溶液取出进行人工定量后再注入 PCR多孔板, 此吋核酸扩增 模块的加热密封盖向外伸出并密封盖合于 PCR多孔板上, 加热密封盖可进行变温 操作, 为 DNA扩增提供合适的反应环境;
[0078] 14. DNA扩增完毕后就能够人工取出, 进行定量处理, 然后再放回系统内进行 后续处理, 加入各种所需试剂, 进行各种反应, 形成所需浓度的 DNA纳米球溶 液;
[0079] 15.形成所需浓度的 DNA纳米球溶液后人工将其取出, 进行定量保存, 最终形 成可以进行基因测序的样本。 [0080] 需要说明的是, 以上步骤仅为本系统的其中一种操作流程, 实验人员可以根据 试验的具体需求设置不同的操作流程。
[0081] 由此可见, 本例的系统可以一站式自动化完成 DNA提取、 酶反应和纯化等建库 操作, 最终获得基因测序样本, 本例具体得到的是 DNA纳米球文库。 本例的系 统可以跟华大基因研发的测序仪配套使用, 为其提供基因测序样本。
[0082] 本例的原材料多孔板可以放置 16个测试样本, 自动移液枪可以同吋装载 8个吸 头, 当然, 还可以根据需要设计成所需数量。
[0083]
[0084] 以上内容是结合具体的实施方式对本申请所作的进一步详细说明, 不能认定本 申请的具体实施只局限于这些说明。 对于本申请所属技术领域的普通技术人员 来说, 在不脱离本申请构思的前提下, 还可以做出若干简单推演或替换, 都应 当视为属于本申请的保护范围。
技术问题
问题的解决方案
发明的有益效果

Claims

权利要求书
[权利要求 1] 一种用于制备基因测序样本的系统, 其特征在于: 包括一个由外壳 (
1) 围成的密封腔体, 以及设置在密封腔体内的原材料放置模块 (2) 、 核酸扩增模块 (3) 、 加热制冷模块 (4) 、 移液模块、 磁珠纯化模 块 (6) 、 废料回收模块和控制模块;
所述外壳 (1) 的顶端幵设有进风口 (11) , 并且, 于进风口 (11) 内安装有空气过滤单元 (12) ;
所述移液模块包括自动移液枪 (51) 和机械臂 (52) , 所述自动移液 枪 (51) 设于所述机械臂 (52) 上, 所述机械臂 (52) 控制所述自动 移液枪 (51) 移动, 所述自动移液枪 (51) 用于吸取和转移液体样本 所述加热制冷模块 (4) 包括电加热组件, 和设置于电加热组件上的 变温多孔板 (41) 、 常温多孔板 (42) , 以及设置于变温多孔板 (41 ) 和常温多孔板 (42) 下方的排气扇 (43) 和散热组件 (44) ; 所述核酸扩增模块 (3) 包括加热密封盖 (31) 、 PCR多孔板 (32) 、 散热器 (33) 和排风扇 (34) , 所述加热密封盖 (31) 在进行 PCR 反应吋将所述 PCR多孔板 (32) 密封, 并进行加热; 所述磁珠纯化模块 (6) 包括纯化多孔板 (61) 、 支架 (62) 、 磁力 架 (63) 和升降机构 (64) , 所述纯化多孔板 (61) 放置于支架 (62 ) 之上, 所述磁力架 (63) 独立的设置于纯化多孔板 (61) 的正下方 , 并且, 所述升降机构 (64) 控制磁力架 (63) 独立上升或下降; 所述废料回收模块包括至少一个废料回收盒 (71) ; 所述控制模块分别与核酸扩增模块 (3) 、 加热制冷模块 (4) 、 移液 模块信号连接, 独立的控制核酸扩增模块 (3) 、 加热制冷模块 (4) 或机械臂 (52) 运行。
[权利要求 2] 根据权利要求 1所述的系统, 其特征在于: 所述外壳 (1) 围成的密封 腔体内还安装有紫外灭菌灯 (9) , 用于对密封腔体进行紫外灭菌消 毒。
[权利要求 3] 根据权利要求 1所述的系统, 其特征在于: 所述外壳 (1) 的前面安装 有一个可幵合的透视安全门 (13) 。
[权利要求 4] 根据权利要求 3所述的系统, 其特征在于: 所述外壳 (1) 的两个侧面 幵设有透视窗口 (14) 。
[权利要求 5] 根据权利要求 4所述的系统, 其特征在于: 所述透视安全门 (13) 和 所述透视窗口 (14) 都采用茶色透明材料制备。
[权利要求 6] 根据权利要求 1所述的系统, 其特征在于: 所述移液模块还包括两个 吸头放置模块 (53、 54) , 用于放置吸头。
[权利要求 7] 根据权利要求 1-6任一项所述的系统, 其特征在于: 所述外壳 (1) 的 背面还设置有 USB转接口 (15) 、 串口控制接口 (16) 、 电源幵关 ( 17) 和网络接口 (18) ; 所述电源幵关 (17) 控制整个系统的电源幵 启或关闭; 所述 USB转接口 (15) 和串口控制接口 (16) 用于连接控 制终端, 包括电脑或其它人机交互界面; 所述网络接口 (18) 用于将 系统与网络连接。
[权利要求 8] 根据权利要求 1-6任一项所述的系统, 其特征在于: 所述外壳 (1) 的 背面还设置有散热口 (19) 。
[权利要求 9] 根据权利要求 1-6任一项所述的系统, 其特征在于: 所述系统还包括 扫码器 (10) , 所述扫码器 (10) 安装于由外壳 (1) 围成的密封腔 体内。
[权利要求 10] 根据权利要求 1-9任一项所述的系统在核酸提取、 核酸纯化或文库构 建中的应用。
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WO2022016475A1 (zh) * 2020-07-23 2022-01-27 深圳华大智造科技股份有限公司 样本前处理系统
CN114134032A (zh) * 2021-12-02 2022-03-04 杭州奥盛仪器有限公司 基因测序前处理装置
WO2023144686A1 (en) * 2022-01-28 2023-08-03 Mylab Discovery Solutions Private Limited A deck assembly for a diagnostic system

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101802164A (zh) * 2007-07-13 2010-08-11 汉迪实验室公司 用于在多个生物样品上进行核酸提取和诊断测试的集成装置
CN104204229A (zh) * 2012-02-10 2014-12-10 株式会社百奥尼 用于自动分析生物试样的设备以及方法
CN205556687U (zh) * 2016-01-20 2016-09-07 广州市基准医疗有限责任公司 核酸测序设备

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100538365C (zh) * 2005-11-01 2009-09-09 清华大学 一种生物芯片点样机器人系统
CN201628714U (zh) * 2010-03-25 2010-11-10 山东博科生物产业有限公司 生物安全生化分析仪
CN203269923U (zh) * 2013-03-15 2013-11-06 北京双诚联盈净化工程技术有限公司 聚合酶链式反应工作站
CN104893951B (zh) * 2014-03-03 2019-03-05 北京双诚联盈净化工程技术有限公司 聚合酶链式反应工作站的组合
CN104089804B (zh) * 2014-07-02 2019-04-05 上海乐辰生物科技有限公司 生物样品处理系统

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101802164A (zh) * 2007-07-13 2010-08-11 汉迪实验室公司 用于在多个生物样品上进行核酸提取和诊断测试的集成装置
CN104204229A (zh) * 2012-02-10 2014-12-10 株式会社百奥尼 用于自动分析生物试样的设备以及方法
CN205556687U (zh) * 2016-01-20 2016-09-07 广州市基准医疗有限责任公司 核酸测序设备

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022016475A1 (zh) * 2020-07-23 2022-01-27 深圳华大智造科技股份有限公司 样本前处理系统
CN114134032A (zh) * 2021-12-02 2022-03-04 杭州奥盛仪器有限公司 基因测序前处理装置
CN114134032B (zh) * 2021-12-02 2022-08-05 杭州奥盛仪器有限公司 基因测序前处理装置
WO2023144686A1 (en) * 2022-01-28 2023-08-03 Mylab Discovery Solutions Private Limited A deck assembly for a diagnostic system

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