CN112251348A - PCR circulating temperature partition control device - Google Patents

PCR circulating temperature partition control device Download PDF

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Publication number
CN112251348A
CN112251348A CN202011145675.7A CN202011145675A CN112251348A CN 112251348 A CN112251348 A CN 112251348A CN 202011145675 A CN202011145675 A CN 202011145675A CN 112251348 A CN112251348 A CN 112251348A
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China
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temperature
temperature control
pcr
stepping motor
platform
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CN202011145675.7A
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CN112251348B (en
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朱飞彪
花昌义
王明龙
吴磊
汪磊
朱灵
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Hefei Institutes of Physical Science of CAS
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Hefei Institutes of Physical Science of CAS
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00Heating or cooling apparatus; Heat insulating devices
    • B01L7/52Heating 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
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/18Means for temperature control
    • B01L2300/1805Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention discloses a PCR circulating temperature partition control device, and relates to the field of molecular biology research and the like. The PCR circulating temperature zone control device comprises a sample placing platform, a sample placing platform support frame, a temperature control platform, a heat insulation plate, a temperature control platform support frame, a stepping motor and a bottom plate. Above-mentioned PCR circulation temperature subregion controlling means when PCR amplification reaction goes on, the motion fast switch-over temperature control district through control step motor provides corresponding reaction temperature for each step of PCR amplification reaction, realizes the fast cycle of PCR biological sample reaction temperature, has saved the time of rising and falling the temperature greatly for the reaction process, improves the amplification efficiency of PCR biological sample, and the device is small, simple structure can satisfy the user demand of different scenes.

Description

PCR circulating temperature partition control device
Technical Field
The invention relates to the field of PCR amplification, in particular to a PCR circulating temperature partition control device
Background
Polymerase Chain Reaction (PCR) is an in vitro DNA amplification technology, and has strong specificity, so it becomes an important support technology in the fields of life science research, clinical molecular diagnosis, food safety detection, environmental microorganism monitoring, etc.
PCR amplification usually undergoes three steps of high temperature denaturation, low temperature annealing (renaturation) and moderate temperature extension for multiple times of cyclic reaction, specific DNA fragments are amplified in large quantities, and a two-step method can also be adopted, namely annealing and extension are carried out simultaneously, so that the one-time temperature rise and fall process is reduced, and the reaction speed is improved.
In the PCR amplification reaction process, firstly, a biological sample is heated to 95 ℃ for high-temperature denaturation reaction, then, the biological sample is cooled to 55 ℃ for low-temperature annealing reaction, and finally, the biological sample is heated to 72 ℃ for moderate-temperature extension reaction, so that a cycle is completed, if a two-step method is adopted, the annealing and the extension in the cycle process can be simultaneously carried out at 60 ℃ to 65 ℃, in the cycle reaction process, the temperature rise and fall process occupies a certain time of the whole reaction process, the PCR amplification process usually needs 30 to 40 cycles or more, and the time spent in the temperature rise and fall process is considerable, so that the detection efficiency of PCR is limited.
Disclosure of Invention
Aiming at the existing problems, the invention provides the PCR circulating temperature partition control device, which can rapidly switch the temperature control area to provide proper reaction conditions for the PCR amplification reaction of the biological sample by controlling the movement of the stepping motor, save the temperature rise and fall time in the whole reaction process and greatly improve the PCR amplification efficiency of the biological sample.
In order to achieve the purpose, the invention adopts the technical scheme that: a PCR cycling temperature partition control device is characterized in that: the device comprises a sample placing platform, a sample placing platform support frame, a temperature control platform, a heat insulation plate, a temperature control platform support frame, a stepping motor and a base;
the sample placing platform comprises a porous plastic net for placing and fixing the PCR tube, and the mesh size is matched with the PCR tube;
the sample placing platform is fixedly connected with the sample platform support frame and is kept horizontal, and the base provides stable support for the sample placing platform;
the temperature control platform is divided into a temperature control area I and a temperature control area II from the middle by a heat insulation plate; the temperature control area I and the temperature control area II are identical in structure and respectively comprise a heat sink, a temperature sensor and a semiconductor heating and refrigerating sheet, a radiator is arranged on the lower surface of the semiconductor heating and refrigerating sheet, and a radiating fan is arranged close to the radiator;
the stepping motor comprises a stepping motor for controlling the temperature control platform to move in the X-axis direction and a stepping motor for controlling the temperature control platform to move in the Y-axis direction; the temperature control platform is fixedly connected with the stepping motor through a temperature control platform support frame, the stepping motor for controlling the X-axis direction movement of the temperature control platform is arranged below and connected with the stepping motor for controlling the Y-axis direction movement of the temperature control platform, and the base provides stable support for the stepping motor and the part above the stepping motor;
and moving the temperature control platform to different temperature control areas through the stepping motor to switch the temperature.
Further, the meshes of the porous plastic net are used for placing and fixing the PCR tube.
Further, the heat insulation plate is a hard hollow double-layer plate.
Further, the temperature control platform is divided into a temperature control area I and a temperature control area II from the middle by a heat insulation plate, and the temperature control area I and the temperature control area II are respectively heated to different preset temperatures.
Furthermore, the temperature control area I and the temperature control area II are used for carrying out PCR amplification two-step method or three-step method experiments.
Furthermore, the step motor comprises a step motor for controlling the temperature control platform to move in the X-axis direction and a step motor for controlling the temperature control platform to move in the Y-axis direction, and the two step motors are connected with the temperature control platform through a support frame.
The heat sink comprises a substrate of metal material for conducting heat to the PCR biological sample, and a plurality of PCR tube placing cavities for placing the PCR biological sample. The substrate made of metal material and the PCR tube placing cavity are made of copper or silver materials with high thermal conductivity and small specific heat capacity, heat can be rapidly conducted to the PCR biological sample, and the reaction precision is improved.
The temperature sensor is embedded in the heat sink, and the temperature sensor monitors and feeds back the temperature of the working surface of the heat sink in real time. The temperature of the temperature control area can be adjusted in real time through data fed back by the temperature sensor.
When the control area needs to be heated, the semiconductor heating and refrigerating piece loads forward voltage through program control, and when refrigeration is needed, reverse voltage is loaded through program control. That is, the rate of temperature increase and decrease can be adjusted by merely controlling and adjusting the direction and magnitude of the current.
The stepping motor and the temperature control platform support frame are connected through four screws, and compression springs are respectively added between the stepping motor and the temperature control platform support frame. The use of compression spring can provide certain buffering again and avoid causing mechanical damage when guaranteeing that PCR pipe places the chamber and the sufficient contact of PCR pipe.
Has the advantages that:
according to the invention, the movement of the stepping motor is controlled, the temperature control area is rapidly switched to provide appropriate reaction conditions for the PCR amplification reaction of the biological sample, the temperature rise and fall time of the whole reaction process is saved, and the PCR amplification efficiency of the biological sample is greatly improved.
Drawings
FIG. 1 is a schematic structural diagram of an embodiment of the present invention;
fig. 2 is a front view of fig. 1.
Wherein: 1-temperature control platform, 10-heat insulation board, 11-temperature control area I, 12-temperature control area II, 111/121-heat sink, 112/122-semiconductor heating and refrigerating sheet, 113/123-radiator, 114/124-cooling fan, 2-temperature control platform support frame, 3-stepping motor for controlling Y-axis direction movement, 4-stepping motor for controlling X-axis direction movement, 5-sample placing platform, 50-PCR tube, 6-sample placing platform support frame and 7-base.
Detailed Description
In order to make those skilled in the art more accurately understand the technical solution of the present invention, the following detailed description and the accompanying drawings are provided to describe and explain the specific embodiments of the present invention in further detail.
As shown in FIG. 1 and FIG. 2, the present invention provides a PCR cycle temperature partition control device, which comprises a sample placement platform 5, a sample placement platform support 6, a temperature control platform 1, a thermal insulation board 10, a temperature control platform support 2, a stepping motor 3/4, and a base 7.
Specifically, as shown in fig. 1 and 2, the sample placement platform 5 comprises a porous plastic net for placing and fixing a PCR tube, the mesh being fitted with the PCR tube 50. Sample placement platform 5 and sample placement platform support frame 6 fixed connection and keep the level, and base 7 provides stable support for the sample platform.
The temperature control platform 1 is divided into a temperature control area I11 and a temperature control area II 12 from the middle by a heat insulation plate 10. Temperature control district I11 and temperature control district II 12 are similar in structure, all include heat sink 111/121, temperature sensor, semiconductor heating refrigeration piece 112/122 respectively, and semiconductor heating refrigeration piece 112/122 lower surface is provided with radiator 113/123, presses close to radiator 113/123 is provided with radiator fan 114/124.
The stepping motor 3/4 includes a stepping motor 4 for controlling the movement of the temperature control stage in the X-axis direction (horizontal direction) and a stepping motor 3 for controlling the movement of the temperature control stage in the Y-axis direction (vertical direction). Temperature control platform 1 is connected fixedly through temperature control platform support frame 2 and step motor 3, and step motor 4 is connected below step motor 3 and with it, and base 6 provides stable support for step motor 4 and step motor top part.
When PCR amplification reaction is carried out, a PCR amplification two-step method is adopted: heating and stabilizing the temperature control area I11 at the denaturation temperature of 93 ℃ in the first step of the PCR amplification reaction, heating and stabilizing the temperature control area II 12 at the renaturation and extension temperatures of 60-65 ℃ in the second step of the PCR amplification reaction, and realizing the rapid temperature switching and the circulation of the PCR amplification reaction only by controlling the stepping motor 3 and the stepping motor 4;
adopting a PCR amplification three-step method: when the temperature control area I11 provides reaction temperature for the PCR biological sample, the temperature control area II 12 adjusts and stabilizes the temperature to the next reaction temperature of the PCR biological sample, after the previous reaction step is finished, the stepping motor 4 is horizontally moved by the descending stepping motor 3, then the stepping motor 3 is ascended to adjust the temperature control area II 12 to the position below the sample placing platform 5 and tightly attach, corresponding temperature is rapidly provided for the next reaction of the PCR biological sample, and by analogy, the circulation of the reaction temperature of the PCR biological sample is realized by rapidly switching the temperature control area. Therefore, no matter which PCR amplification mode is adopted, the device can greatly save the time for heating and cooling, and improve the amplification efficiency of the PCR biological sample.
The present invention has been described in detail with reference to the accompanying drawings. It will be appreciated by persons skilled in the art that the foregoing is merely illustrative and that specific implementations of the invention are not limited by the foregoing embodiments. Insubstantial changes from the embodiments are intended to be within the scope of the invention.

Claims (10)

1. A PCR cycling temperature partition control device is characterized in that: the device comprises a sample placing platform, a sample placing platform support frame, a temperature control platform, a heat insulation plate, a temperature control platform support frame, a stepping motor and a base;
the sample placing platform comprises a porous plastic net for placing and fixing the PCR tube, and the mesh size is matched with the PCR tube;
the sample placing platform is fixedly connected with the sample platform support frame and is kept horizontal, and the base provides stable support for the sample placing platform;
the temperature control platform is divided into a temperature control area I and a temperature control area II from the middle by a heat insulation plate; the temperature control area I and the temperature control area II are identical in structure and respectively comprise a heat sink, a temperature sensor and a semiconductor heating and refrigerating sheet, a radiator is arranged on the lower surface of the semiconductor heating and refrigerating sheet, and a radiating fan is arranged close to the radiator;
the stepping motor comprises a stepping motor for controlling the temperature control platform to move in the X-axis direction and a stepping motor for controlling the temperature control platform to move in the Y-axis direction; the temperature control platform is fixedly connected with the stepping motor through a temperature control platform support frame, the stepping motor for controlling the movement in the X-axis direction is arranged below and connected with the stepping motor for controlling the movement in the Y-axis direction, and the base provides stable support for the stepping motor and the part above the stepping motor;
and moving the temperature control platform to different temperature control areas through the stepping motor to switch the temperature.
2. The PCR cycling temperature partition control device according to claim 1, characterized in that: the mesh of the porous plastic net is used for placing and fixing the PCR tube.
3. The PCR cycling temperature partition control device according to claim 1, characterized in that: the heat insulation plate is a hard hollow double-layer plate.
4. The PCR cycling temperature partition control device according to claim 1, characterized in that: the temperature control platform is divided into a temperature control area I and a temperature control area II from the middle by a heat insulation plate, and the temperature control area I and the temperature control area II are respectively heated to different preset temperatures.
5. The PCR cycling temperature partition control device according to claim 4, characterized in that: the temperature control area 1 and the temperature control area II are used for carrying out PCR amplification two-step method or three-step method experiments.
6. The PCR cycling temperature partition control device according to claim 1, characterized in that: the heat sink includes a substrate of a metal material for conducting heat to the PCR biological samples, and a plurality of PCR tube placement chambers for placing the PCR biological samples.
7. The PCR cycling temperature partition control device according to claim 1, characterized in that: the temperature sensor is pre-embedded in the heat sink, and carries out real-time monitoring and feedback on the temperature of the working surface of the heat sink.
8. The PCR cycling temperature partition control device according to claim 6, characterized in that: in the working process of the semiconductor heating and refrigerating piece, when the control area needs to be heated, forward voltage is loaded through program control, and when refrigeration is needed, reverse voltage is loaded through program control.
9. The PCR cycling temperature partition control device according to claim 1, characterized in that: the stepping motors comprise a stepping motor for controlling the temperature control platform to move in the X-axis direction and a stepping motor for controlling the temperature control platform to move in the Y-axis direction, and the two stepping motors are connected with the temperature control platform through a support frame.
10. The PCR cycling temperature partition control device according to claim 1, characterized in that: the stepping motor and the temperature control platform support frame are connected through four screws, and compression springs are respectively added between the stepping motor and the temperature control platform support frame.
CN202011145675.7A 2020-10-23 2020-10-23 PCR circulation temperature partition control device Active CN112251348B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114134037A (en) * 2021-12-17 2022-03-04 新羿制造科技(北京)有限公司 PCR amplification temperature control device

Citations (5)

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Publication number Priority date Publication date Assignee Title
CN204939449U (en) * 2015-06-09 2016-01-06 吕明月 The portable PCR instrument of a kind of new microization
CN107574120A (en) * 2017-10-27 2018-01-12 深圳大学 Fluorescence quantitative PCR detection system and method based on magnetomotive switching flat-temperature zone
CN110387325A (en) * 2019-08-13 2019-10-29 广州市华南医学研究中心 A kind of very fast PCR reaction checking device and detection method
CN110564610A (en) * 2019-10-15 2019-12-13 杭州比芯诊断技术有限公司 Double-temperature-zone PCR amplification device
CN210215375U (en) * 2019-04-13 2020-03-31 浙江东吉生物科技有限公司 PCR fluorescence detector

Patent Citations (5)

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Publication number Priority date Publication date Assignee Title
CN204939449U (en) * 2015-06-09 2016-01-06 吕明月 The portable PCR instrument of a kind of new microization
CN107574120A (en) * 2017-10-27 2018-01-12 深圳大学 Fluorescence quantitative PCR detection system and method based on magnetomotive switching flat-temperature zone
CN210215375U (en) * 2019-04-13 2020-03-31 浙江东吉生物科技有限公司 PCR fluorescence detector
CN110387325A (en) * 2019-08-13 2019-10-29 广州市华南医学研究中心 A kind of very fast PCR reaction checking device and detection method
CN110564610A (en) * 2019-10-15 2019-12-13 杭州比芯诊断技术有限公司 Double-temperature-zone PCR amplification device

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Title
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114134037A (en) * 2021-12-17 2022-03-04 新羿制造科技(北京)有限公司 PCR amplification temperature control device
CN114134037B (en) * 2021-12-17 2023-12-29 新羿制造科技(北京)有限公司 PCR amplification temperature control device

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