CN112251348B - PCR circulation temperature partition control device - Google Patents
PCR circulation temperature partition control device Download PDFInfo
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- CN112251348B CN112251348B CN202011145675.7A CN202011145675A CN112251348B CN 112251348 B CN112251348 B CN 112251348B CN 202011145675 A CN202011145675 A CN 202011145675A CN 112251348 B CN112251348 B CN 112251348B
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- temperature control
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- stepping motor
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- 238000005192 partition Methods 0.000 title claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 33
- 239000000523 sample Substances 0.000 claims abstract description 24
- 239000012472 biological sample Substances 0.000 claims abstract description 23
- 238000012408 PCR amplification Methods 0.000 claims abstract description 19
- 238000009413 insulation Methods 0.000 claims abstract description 11
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000004065 semiconductor Substances 0.000 claims description 10
- 230000006835 compression Effects 0.000 claims description 3
- 238000007906 compression Methods 0.000 claims description 3
- 239000007769 metal material Substances 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 2
- 230000001174 ascending effect Effects 0.000 claims 1
- 238000005057 refrigeration Methods 0.000 claims 1
- 230000003321 amplification Effects 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 230000000630 rising effect Effects 0.000 abstract 1
- 238000000137 annealing Methods 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 3
- 230000036425 denaturation Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 238000004153 renaturation Methods 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000017525 heat dissipation Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
Classifications
-
- 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
-
- 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
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
Landscapes
- 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 circulation temperature partition control device, and relates to the fields of molecular biology research and the like. The PCR circulation temperature partition control device comprises a sample placement platform, a sample placement platform support frame, a temperature control platform, a heat insulation plate, a temperature control platform support frame, a stepping motor and a bottom plate. According to the PCR circulation temperature partition control device, when the PCR amplification reaction is carried out, the corresponding reaction temperature is provided for each step of the PCR amplification reaction by controlling the movement quick switching temperature control area of the stepping motor, so that the quick circulation of the reaction temperature of the PCR biological sample is realized, the temperature rising and reducing time is greatly saved, the reaction process is accelerated, the amplification efficiency of the PCR biological sample is improved, the device is small in size and simple in structure, and the use requirements of different scenes can be met.
Description
Technical Field
The invention relates to the field of PCR amplification, in particular to a PCR circulation temperature partition control device
Background
The polymerase chain reaction (Polymerase Chain Reaction, PCR) is an in vitro DNA amplification technology, and has strong specificity, so that the PCR is an important support technology in the fields of life science research, clinical molecular diagnosis, food safety detection, environmental microorganism monitoring and the like.
The PCR amplification usually adopts three steps of high temperature denaturation, low temperature annealing (renaturation) and moderate temperature extension to carry out repeated cyclic reaction, and then a specific DNA fragment is greatly amplified, and a two-step method, namely annealing and extension are carried out simultaneously, so that the one-time temperature raising and lowering process is reduced, and the reaction speed is improved.
In the PCR amplification reaction process, firstly, a biological sample is heated to 95 ℃ to carry out high-temperature denaturation reaction, then the biological sample is cooled to 55 ℃ to carry out low-temperature annealing reaction, finally, the biological sample is heated to 72 ℃ to carry out temperature-adaptive extension reaction, thus completing a cycle, if a two-step method is adopted, annealing and extension in the cycle process are carried out at 60-65 ℃ simultaneously, in the cycle reaction process, the temperature-increasing process occupies a certain time in the whole reaction process, the PCR amplification process usually needs to pass through 30-40 cycles or more, the time spent in the temperature-increasing process is considerable, and thus the detection efficiency of PCR is limited.
Disclosure of Invention
Aiming at the problems, the invention provides the PCR circulation temperature partition control device, which provides proper reaction conditions for the PCR amplification reaction of the biological sample by controlling the movement of the stepping motor and rapidly switching the temperature control area, thereby saving the temperature rise and fall time in the whole reaction process and greatly improving the PCR amplification efficiency of the biological sample.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: a PCR circulation temperature partition control device is characterized in that: the device comprises a sample placing platform, a sample placing platform supporting frame, a temperature control platform, a heat insulation plate, a temperature control platform supporting 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 supporting frame and 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 have the same structure and respectively comprise a heat sink, a temperature sensor and a semiconductor heating and refrigerating sheet, wherein the lower surface of the semiconductor heating and refrigerating sheet is provided with a radiator, 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 of the temperature control platform in the X-axis direction is arranged below and connected with the stepping motor for controlling the movement of the temperature control platform in the Y-axis direction, and the base provides stable support for the stepping motor and the part above the stepping motor;
and (5) moving the temperature control platform to different temperature control areas through a stepping motor to perform temperature switching.
Further, the mesh of the porous plastic net is used for placing and fixing the PCR tube.
Further, the heat insulation board is a hard hollow double-layer board.
Further, the temperature control platform is divided into a temperature control area I and a temperature control area II from the middle by the 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 region I and the temperature control region II are used for performing a two-step or three-step PCR amplification experiment.
Further, the stepper motor comprises a stepper motor for controlling the X-axis direction of the temperature control platform to move and a stepper motor for controlling the Y-axis direction of the temperature control platform to move, and the two stepper motors are connected with the temperature control platform through a supporting frame.
The heat sink comprises a substrate made 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 of the metal material and the PCR tube placing cavity are made of copper or silver materials with high heat conductivity and small specific heat capacity, so that heat can be quickly conducted to a PCR biological sample, and the reaction accuracy is improved.
The temperature sensor is pre-buried in the heat sink, and the temperature sensor monitors and feeds back the temperature of the working face of the heat sink in real time. The temperature of the temperature control area can be adjusted in real time through the data fed back by the temperature sensor.
When the control area needs to be heated, the semiconductor heating and refrigerating sheet loads forward voltage through program control, and when the semiconductor heating and refrigerating sheet needs to be refrigerated, reverse voltage is loaded through program control. That is, the rate of temperature rise and fall can be adjusted by 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 arranged between the stepping motor and the temperature control platform support frame. The use of the compression spring can ensure that the PCR tube placing cavity is fully contacted with the PCR tube and can provide certain buffer to avoid mechanical damage.
The beneficial effects are that:
the invention provides proper reaction conditions for the PCR amplification reaction of the biological sample by controlling the movement of the stepping motor and rapidly switching the temperature control area, thereby saving the temperature rise time of the whole reaction process and greatly improving the PCR amplification efficiency of the biological sample.
Drawings
FIG. 1 is a schematic diagram of an embodiment of the present invention;
fig. 2 is a front view of fig. 1.
Wherein: the device comprises a 1-temperature control platform, a 10-heat insulation plate, a 11-temperature control area I, a 12-temperature control area II, a 111/121-heat sink, a 112/122-semiconductor heating and refrigerating sheet, a 113/123-heat radiator, a 114/124-heat radiation fan, a 2-temperature control platform support frame, a 3-stepper motor for controlling Y-axis direction movement, a 4-stepper motor for controlling X-axis direction movement, a 5-sample placement platform, a 50-PCR tube, a 6-sample placement platform support frame and a 7-base.
Detailed Description
In order to enable those skilled in the art to understand the technical scheme of the present invention more accurately, the following detailed description and the description of the specific embodiments of the present invention are provided with reference to the accompanying drawings.
As shown in fig. 1 and 2, the 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 heat insulation board 10, a temperature control platform support 2, a stepping motor 3/4 and a base 7.
In particular, as shown in fig. 1 and 2, the sample placement platform 5 comprises a porous plastic mesh for placement and immobilization of PCR tubes, the mesh being adapted to PCR tubes 50. The sample placement platform 5 is fixedly connected with the sample placement platform support frame 6 and kept horizontal, and the base 7 provides stable support for the sample placement platform.
The temperature control platform 1 is divided into a temperature control zone I11 and a temperature control zone II 12 from the middle by a heat insulating plate 10. The temperature control area I11 and the temperature control area II 12 are similar in structure and respectively comprise a heat sink 111/121, a temperature sensor and a semiconductor heating and refrigerating sheet 112/122, a radiator 113/123 is arranged on the lower surface of the semiconductor heating and refrigerating sheet 112/122, and a heat dissipation fan 114/124 is arranged close to the radiator 113/123.
The stepper motors 3/4 include a stepper motor 4 that controls the movement of the temperature control platform in the X-axis direction (horizontal direction) and a stepper motor 3 that controls the movement of the temperature control platform in the Y-axis direction (vertical direction). The temperature control platform 1 is fixedly connected with the stepper motor 3 through the temperature control platform support frame 2, the stepper motor 4 is arranged below the stepper motor 3 and connected with the stepper motor, and the base 6 provides stable support for the stepper motor 4 and the part above the stepper motor.
In the PCR amplification reaction, a two-step PCR amplification method is adopted: the temperature control area I11 is heated and stabilized at the denaturation temperature of 93 ℃ in the first step of the PCR amplification reaction, the temperature control area II 12 is heated and stabilized at the renaturation and extension temperature of 60 ℃ to 65 ℃ in the second step of the PCR amplification reaction, and the rapid temperature switching and the circulation of the PCR amplification reaction can be realized only by controlling the stepper motor 3 and the stepper motor 4;
the three-step PCR amplification method comprises the following steps: when the temperature control area I11 provides the 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 last reaction step is finished, the stepping motor 3 is moved horizontally by the descending stepping motor 4 and then the stepping motor 3 is lifted up to adjust the temperature control area II 12 to the lower part of the sample placing platform 5 and is tightly attached, the corresponding temperature is provided for the next reaction of the PCR biological sample rapidly, and 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 of temperature rise and reduction and improve the amplification efficiency of the PCR biological sample.
The foregoing is a detailed description of the invention with reference to the accompanying drawings. It will be appreciated by those skilled in the art that the foregoing is by way of example only and that the invention is not limited to the embodiments described. Insubstantial changes in the embodiments are still within the scope of the invention.
Claims (6)
1. A PCR circulation temperature partition control device is characterized in that: the device comprises a sample placing platform, a sample placing platform supporting frame, a temperature control platform, a heat insulation plate, a temperature control platform supporting 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 supporting frame and 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 have the same structure and respectively comprise a heat sink, a temperature sensor and a semiconductor heating and refrigerating sheet, wherein the lower surface of the semiconductor heating and refrigerating sheet is provided with a radiator, 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 which controls the movement of the X axis direction is arranged below the stepping motor which controls the movement of the Y axis direction and is connected with the stepping motor, and the base provides stable support for the stepping motor and the part above the stepping motor;
the temperature control platform is moved to different temperature control areas through the stepping motor to switch the temperature;
the meshes of the porous plastic net are used for placing and fixing the PCR tube;
the heat insulation plate is a hard hollow double-layer plate;
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 is heated to different preset temperatures respectively;
the temperature control area 1 and the temperature control area II are used for performing a two-step or three-step PCR amplification experiment;
when the temperature control area I provides a reaction temperature for the PCR biological sample, the temperature control area II adjusts and stabilizes the temperature to the next reaction temperature of the PCR biological sample, after the last reaction step is finished, the temperature control area II is adjusted to the lower part of the sample placing platform and is tightly attached by the descending stepping motor and the ascending stepping motor, so that the corresponding temperature is rapidly provided for the next reaction of the PCR biological sample, and the circulation of the reaction temperature of the PCR biological sample is realized by rapidly switching the temperature control area.
2. The PCR cycle temperature zone control device as claimed in claim 1, wherein: the heat sink includes a substrate of a metal material for conducting heat to the PCR biological sample, and a plurality of PCR tube placement chambers for placing the PCR biological sample.
3. The PCR cycle temperature zone control device as claimed in claim 1, wherein: the temperature sensor is pre-buried in the heat sink, and monitors and feeds back the temperature of the working face of the heat sink in real time.
4. A PCR cycle temperature zone control device in accordance with claim 3, wherein: in the working process of the semiconductor heating and refrigerating sheet, 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.
5. The PCR cycle temperature zone control device as claimed in claim 1, wherein: 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, and the two stepping motors are connected with the temperature control platform through a supporting frame.
6. The PCR cycle temperature zone control device as claimed in claim 1, wherein: the stepping motor and the temperature control platform support frame are connected through four screws, and compression springs are respectively arranged between the stepping motor and the temperature control platform support frame.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011145675.7A CN112251348B (en) | 2020-10-23 | 2020-10-23 | PCR circulation temperature partition control device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011145675.7A CN112251348B (en) | 2020-10-23 | 2020-10-23 | PCR circulation temperature partition control device |
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| Publication Number | Publication Date |
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| CN112251348A CN112251348A (en) | 2021-01-22 |
| CN112251348B true CN112251348B (en) | 2024-03-26 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202011145675.7A Active CN112251348B (en) | 2020-10-23 | 2020-10-23 | PCR circulation temperature partition control device |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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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|---|---|---|---|---|
| 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 |
-
2020
- 2020-10-23 CN CN202011145675.7A patent/CN112251348B/en active Active
Patent Citations (5)
| 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 |
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| CN112251348A (en) | 2021-01-22 |
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