CN109971617A - A kind of cryogenic process system of PCR amplification device - Google Patents
A kind of cryogenic process system of PCR amplification device Download PDFInfo
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- CN109971617A CN109971617A CN201910361197.4A CN201910361197A CN109971617A CN 109971617 A CN109971617 A CN 109971617A CN 201910361197 A CN201910361197 A CN 201910361197A CN 109971617 A CN109971617 A CN 109971617A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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Abstract
The invention discloses a kind of cryogenic process system of PCR amplification device and PCR amplification devices, it include: the circulation loop for cryogen circulation for being connected to and being formed by low temperature control module, the first flow velocity control module and reaction unit, circulation loop maintains the reaction temperature in reaction unit to be in constant low temperature reaction interval；Low temperature control module is for maintaining and exporting constant cryogen；First flow velocity control module is by improving the flow velocity of cryogen, so that greatly shortening reaction unit reaches the time required for low-temp reaction section.The circulation loop for high temperature fluid circulation formed is connected to by high-temperature control module, second flow speed control module and reaction unit, circulation loop maintains the reaction temperature in reaction unit to be in constant high temperature reaction interval；High-temperature control module is for maintaining and exporting constant high temperature fluid；Second flow speed control module is by promoting the flow velocity of high temperature fluid, so that greatly shortening reaction unit reaches the time required for pyroreaction section.
Description
Technical field
The present invention relates to PCR amplification device fields, more particularly, to the PCR amplification that can reach low-temp reaction temperature rapidly
Device.
Background technique
With the development of bioscience technology and genetic engineering, DNA(DNA) research in field becomes current
The hot spot of biological study.Become single-stranded at high temperature according to DNA, what when low temperature combined according to the principle of complementary pairing
Principle.PCR(polymerase chain reaction) amplification device comes into being, and it passes through the ginseng that needs during artificially simulation PCR amplification
PCR amplification process is divided into the thermal cycle of three high-temperature denaturation, low-temperature annealing and appropriate temperature extension steps repeatedly and formed by number.
Scientific investigations showed that reaction temperature and reaction time have a major impact the expanding effect of PCR, in reaction temperature
Closer to theoretical value, the reaction time is rapider for low-temp reaction section and pyroreaction section, and obtained experimental result is more valuable.
But current existing PCR amplification device, it is directed generally to improve the pyroprocess during PCR amplification, few devices pair
Chilling process and shortening reaction time during PCR amplification are improved.However the reaction temperature of low-temp reaction process and anti-
The complementary fit stage in PCR amplification is directly determined between seasonable.Reaction temperature and the improper of reaction time will lead to PCR expansion
Difficulty occurs for the complementary fit in increasing process, influences PCR amplification effect.
Summary of the invention
For the technological deficiency more than solving, it is an object of that present invention to provide a kind of low-temperature treatment systems of PCR amplification device
System and PCR amplification device, the PCR amplification device include cryogenic process system and high-temperature process system, the high-temperature process
System and cryogenic process system share a reaction unit, moving back property of low temperature and high-temperature denaturation alternately during PCR amplification
Stage.
Specifically, the reaction unit includes pedestal, reservoir and insulation construction；The reservoir and insulation construction peace
Loaded on the base interior.The reservoir is used for transmission fluid, and the insulation construction is used for placing response reagent.
The cryogenic process system includes:
Low temperature control module, the first flow velocity control module and reaction unit are connected to being recycled back to for cryogen circulation to be formed
Road；The circulation loop maintains the reaction temperature to be in constant low temperature reaction interval；The low temperature control module generates temperature
The constant cryogen is to be conveyed to the first flow velocity control module；The first flow velocity control module is promoted described low
The flow velocity of warm fluid is rapidly reached the low-temp reaction section to control the reaction temperature.
Wherein, the low temperature control module includes the first liquid storage device for storing and exporting constant cryogen, low temperature
Controller is for maintaining fluid constant low temperature；First flow velocity control module includes low temperature into pumping and move back pump unit, low temperature into pump and
Pump unit is moved back by improving the flow velocity of the cryogen, shorten the reaction temperature reach the low-temp reaction section when
Between.
Specifically, first liquid storage device uses vacuum heat-insulating layer, guarantees that liquid storage device internal temperature and external temperature are separated by
From the first liquid storage device internal temperature constant low temperature of maintenance.
Specifically, the cryostat is reduced in the first liquid storage device using chemical reaction falling temperature method or automatically controlled preparation method
The fluid temperature (F.T.) in portion guarantees that low temperature control module continues to export cryogen outward.
Specifically, the low temperature in the first flow velocity control module into pump and low temperature move back pump unit, be set as positive displacement or
Person's vane type oil pump.The low temperature moves back pump unit by changing flow velocity of the fluid in circulation loop into pump unit and low temperature, changes
Reaction time in reaction unit.
Specifically, switch is added in the first flow velocity control module, and switch is set as mechanical valve or solenoid valve, to described
Circulation loop carries out switch control.When the switch is opened, the PCR amplification device opens cryogenic process system, and experiment enters
Low temperature moves back the sexual stage.
Specifically, the low temperature moves back the sexual stage, and the reservoir is constantly constant low to inputting inside the insulation construction
Warm fluid maintains the temperature in reaction unit to be in constant low temperature.
The high-temperature process system includes:
High-temperature control module, second flow speed control module and reaction unit are connected to being recycled back to for high temperature fluid circulation to be formed
Road；The circulation loop maintains the reaction temperature to be in constant high temperature reaction interval.
Specifically, the high-temperature control module generates the constant high temperature fluid of temperature to be conveyed to the second flow speed
Control module；The second flow speed control module promotes the flow velocity of the high temperature fluid and is rapidly reached with controlling the reaction temperature
The pyroreaction section.
Specifically, the high-temperature control module includes the second liquid storage device and pyrostat, is respectively used to store constant
High temperature fluid and maintenance fluid constant high temperature；The second flow speed control module includes high temperature into pumping and move back pump unit, the height
Temperature moves back pump unit by improving the flow velocity of the high temperature fluid into pump and high temperature, and shortening the reaction temperature, to reach the high temperature anti-
Answer the time in section.
Specifically, second liquid storage device uses vacuum heat-insulating layer, guarantees that liquid storage device internal temperature and external temperature are separated by
From to maintain the second liquid storage device internal temperature constant high temperature.
Specifically, the pyrostat is using chemical reaction temperature-raising method, Electric control method or physical heating method, heating
Fluid temperature (F.T.) inside second liquid storage device guarantees that high-temperature control module continues to export high temperature fluid outward.
Specifically, the high temperature in the second flow speed control module into pump and high temperature move back pump unit, be set as positive displacement or
Person's vane type oil pump.High temperature moves back pump unit by changing flow velocity of the fluid in circulation loop, to change into pump unit and high temperature
Reaction time in reaction unit.
Specifically, switch is added in the second flow speed control module, carries out switch control to the circulation loop.Described
When the switch of two flow velocity control modules is opened, the PCR amplification device opens high-temperature process system, and experiment enters high-temperature denaturation rank
Section.
Specifically, the high-temperature denaturation stage, the reservoir are constantly inputting constant high temperature stream to the insulation construction
Body maintains the temperature in reaction unit to be in constant high temperature state.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for the PCR amplification device that embodiment provides.
Fig. 2 is the structural schematic diagram of the cryogenic process system for the PCR amplification device that embodiment provides.
Fig. 3 is the structural schematic diagram of the high-temperature process system for the PCR amplification device that embodiment provides.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
Technical solution in the embodiment of the present invention carries out clear and complete description；Obviously, described embodiment is only the present invention one
Section Example, instead of all the embodiments；Based on the embodiment of the present invention, those of ordinary skill in the art are not making wound
Every other embodiment obtained under the premise of the property made labour, shall fall within the protection scope of the present invention.
Embodiment:
Refering to fig. 1, the present invention provides a kind of technical solution: the PCR amplification device includes cryogenic process system as described in Figure 2
With high-temperature process system as described in Figure 3；Cryogenic process system as described in Figure 2 and high-temperature process system alternating as described in Figure 3
Work, the two share reaction unit 2, while by the control of master control system 1；The master control system is by computer equipment, software
Equipment and data line 15 form, and the software equipment acquires the cryogenic process system and high-temperature process system by data line
In Parameters variation, the experimental state during PCR amplification is controlled.
Cryogenic process system as described in Figure 2 include by low temperature control module 17, the first flow velocity control module 18 and
Reaction unit 2 is connected to the circulation loop for cryogen circulation of formation；The low temperature control module 17 includes: the first liquid storage
Device 3, cryostat 5 and transfer tube 16；The cryostat 5 is mounted on inside the first liquid storage device 3, is made using thermocouple
For temperature sensing device, temperature signal is transformed into electric signal, feeds back to master control system, so that fluid temperature (F.T.) is maintained 50 DEG C
To between 55 DEG C；First liquid storage device 3 uses stainless steel material, in the 3 outer packaging thermal insulation layer of the first liquid storage device, thus
Reach and maintains cryogen between 50 DEG C to 55 DEG C；The first flow velocity control module 18 includes switch 8, low temperature into pump unit
7, switch 10 and low temperature move back pump unit 9；The switch 8 and switch 10 use electromagnetic valve switch, described for starting and stopping
Cryogenic process system；Low temperature moves back pump unit 9 into pump unit 7 and low temperature and uses positive displacement pump, low in transfer tube 16 for adjusting
Warm fluid flow rate.
High-temperature process system as described in Figure 3 include by high-temperature control module 19, second flow speed control module 20 and
Reaction unit 2 is connected to the circulation loop for high temperature fluid circulation of formation；The high-temperature control module 19 includes: the second liquid storage
Device 4, pyrostat 6 and transfer tube 16；The pyrostat 6 is mounted on inside the second liquid storage device 4, using heating wire plus
Temperature signal is transformed into electric signal, feeds back to master control system by hot mode using thermocouple as temperature sensing device, thus
Fluid temperature (F.T.) is maintained between 90 DEG C to 95 DEG C；Second liquid storage device 4 uses stainless steel material, in second liquid storage device
4 installation thermal insulation layers, to keep fluid constant between 90 DEG C to 95 DEG C；The second flow speed control module 20 includes switch
12, into pump unit 11, switch 14 and pump unit 13 is moved back；The switch 12 and switch 14 use electromagnetic valve switch, for starting
With the stopping high-temperature process system；High temperature moves back pump unit 13 using positive displacement pump into pump unit 11 and low temperature, passes for adjusting
High temperature fluid flow velocity in defeated pipe 16.
When PCR amplification device is worked, according to PCR amplification principle, the low temperature for being divided into DNA moves back the sexual stage and high temperature becomes
Sexual stage.
It is moved back the sexual stage when in low temperature, the cryostat 5 in the low temperature control module 17 starts to cool down, by the first storage
Fluid in liquid device 3 is reduced to 50 DEG C to 55 DEG C, by operating master control system 1, opens in the first flow velocity control module 18
Switch 8, into pump unit 7, switch 10 and move back pump unit 9；At this time low temperature control module 17, the first flow velocity control module 18 with
And the connection of reaction unit 2 forms closed circuit, arrow direction of the cryogen in figure one is flowed.In this circuit, begin
The fluid for keeping low temperature control module 17 to export eventually flows through reaction dress by improving in low-temperature condition, the first flow velocity control module 18
The flow velocity of the cryogen in 2 is set, to shorten the time that reaction temperature is reduced to low-temp reaction section rapidly, accelerates low temperature and moves back
The reaction efficiency of sexual stage.
When being in the high-temperature denaturation stage, the pyrostat 6 in the high-temperature control module 19 is started to warm up, by the second storage
Fluid in liquid device 4 is increased to 90 DEG C to 95 DEG C, by operating master control system 1, opens in the second flow speed control module 20
Switch 12, into pump unit 11, switch 14 and move back pump unit 13；High-temperature control module 19, second flow speed control module at this time
20 and reaction unit 2 connection form closed circuit.In this circuit, the fluid stream that high-temperature control module 19 exports is remained
Body is maintained at the condition of high temperature, and second flow speed control module 20 flows through the flow velocity of the high temperature fluid in reaction unit 2 by improving,
To shorten the time that reaction temperature is increased to pyroreaction section rapidly, the reaction efficiency in high-temperature denaturation stage is promoted.
The above is only presently preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Mind and principle within made any modification, equally mention and change and improve, should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of cryogenic process system of PCR amplification device characterized by comprising pass through low temperature control module, the first flow velocity
Control module and reaction unit are connected to the circulation loop for cryogen circulation to be formed；The circulation loop maintains the reaction
Temperature is in constant low temperature reaction interval；The low temperature control module generates the constant cryogen of temperature to be conveyed to
State the first flow velocity control module；The first flow velocity control module promotes the flow velocity of the cryogen to control the reaction temperature
Degree is rapidly reached the low-temp reaction section.
2. the cryogenic process system of PCR amplification device as described in the appended claim 1, it is characterised in that: the low temperature control module
Including the first liquid storage device and cryostat；The cryostat controls the cryogen of the first liquid storage device storage
Maintain constant low temperature；First liquid storage device exports the constant cryogen.
3. the cryogenic process system of PCR amplification device as described in the appended claim 1, it is characterised in that: first flow control
Module includes low temperature governor；It is rapid to control the reaction temperature that the low temperature governor accelerates the flow velocity of the cryogen
Reach the low-temp reaction section.
4. the cryogenic process system of PCR amplification device as claimed in claim 3, it is characterised in that: the low temperature governor packet
It includes low temperature and is installed on the input terminal of the reaction unit into pump unit into pumping and moving back pump unit, the low temperature, the low temperature moves back pump
Unit is installed on the output end of the reaction unit；The low temperature is into pump and the pump unit of moving back by improving the cryogen
Flow velocity, shorten the time that the reaction temperature reaches the low-temp reaction section.
5. a kind of PCR amplification device characterized by comprising such as the described in any item cryogenic process systems of claims 1 to 4.
6. PCR amplification device as claimed in claim 5 further includes high-temperature process system；The high-temperature process system and described
Cryogenic process system works alternatively；The high-temperature process system includes high-temperature control module, second flow speed control module and described
Reaction unit is connected to the circulation loop for high temperature fluid circulation to be formed；The circulation loop maintains the reaction temperature to be in perseverance
Determine pyroreaction section；The high-temperature control module generates the constant high temperature fluid of temperature to be conveyed to the second flow speed
Control module；The second flow speed control module promotes the flow velocity of the high temperature fluid and is rapidly reached with controlling the reaction temperature
The pyroreaction section.
7. PCR amplification device as recited in claim 6, it is characterised in that: the high-temperature control module includes the second liquid storage device
And pyrostat；The high temperature fluid that the pyrostat controls the second liquid storage device storage maintains constant high temperature；
Second liquid storage device exports the constant high temperature fluid.
8. PCR amplification device as recited in claim 6, it is characterised in that: the second flow speed control module includes high temperature tune
Fast device；The high temperature governor accelerates the flow velocity of the high temperature fluid, and to control the reaction temperature, to be rapidly reached the high temperature anti-
Answer section.
9. PCR amplification device as claimed in claim 8, it is characterised in that: the high temperature governor includes high temperature into pump and high
Temperature moves back pump unit, and the high temperature is installed on the input terminal of the reaction unit into pump unit, and the high temperature moves back pump unit and is installed on
The output end of the reaction unit；High temperature moves back pump unit by the flow velocity of the quickening high temperature fluid into pump and high temperature, shortens institute
State the time that reaction temperature reaches the pyroreaction section.
10. PCR amplification device as recited in claim 6, it is characterised in that: the high temperature and low temperature have relativity；It is described
High temperature refers to that the low temperature refers to lower than 80 DEG C higher than 80 DEG C；The pyroreaction section is 90 DEG C to 95 DEG C；The cryogenic reaction zone
Between be 50 DEG C to 60 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201910361197.4A CN109971617A (en) | 2019-04-30 | 2019-04-30 | A kind of cryogenic process system of PCR amplification device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910361197.4A CN109971617A (en) | 2019-04-30 | 2019-04-30 | A kind of cryogenic process system of PCR amplification device |
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| CN201910361197.4A Pending CN109971617A (en) | 2019-04-30 | 2019-04-30 | A kind of cryogenic process system of PCR amplification device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113201454A (en) * | 2021-05-25 | 2021-08-03 | 江南大学 | Small-size nucleic acid rapid annealing device using liquid exchange method |
| CN114608353A (en) * | 2022-03-04 | 2022-06-10 | 通用生物(滁州)有限公司 | Temperature compensation system of oligonucleotide production reaction system |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0402995A2 (en) * | 1989-06-12 | 1990-12-19 | Johnson & Johnson Clinical Diagnostics, Inc. | Temperature control device and reaction vessel |
| CN2767454Y (en) * | 2004-07-06 | 2006-03-29 | 北京工业大学 | Application package structure for microfluidic chip for PCR amplification |
| JP2009517075A (en) * | 2005-11-30 | 2009-04-30 | デルタドット・リミテッド | Real-time PCR monitoring using intrinsic imaging without labeling |
| CN102046291A (en) * | 2008-04-04 | 2011-05-04 | It-Is国际有限公司 | Thermal control system and method for chemical and biochemical reactions |
| CN102559488A (en) * | 2012-01-16 | 2012-07-11 | 福建医科大学 | Quantitative polymerase chain reaction (PCR) microfluidic chip integrated device for integrated electrochemical detection technology |
| WO2016006612A1 (en) * | 2014-07-08 | 2016-01-14 | 国立研究開発法人産業技術総合研究所 | Nucleic acid amplification device, nucleic acid amplification method, and chip for nucleic acid amplification |
| CN106052198A (en) * | 2016-05-31 | 2016-10-26 | 大连海事大学 | PCR amplifier with fast temperature raising and lowering function |
| WO2017199933A1 (en) * | 2016-05-18 | 2017-11-23 | 日本板硝子株式会社 | Reaction treatment device, and method for controlling reaction treatment device |
-
2019
- 2019-04-30 CN CN201910361197.4A patent/CN109971617A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0402995A2 (en) * | 1989-06-12 | 1990-12-19 | Johnson & Johnson Clinical Diagnostics, Inc. | Temperature control device and reaction vessel |
| CN2767454Y (en) * | 2004-07-06 | 2006-03-29 | 北京工业大学 | Application package structure for microfluidic chip for PCR amplification |
| JP2009517075A (en) * | 2005-11-30 | 2009-04-30 | デルタドット・リミテッド | Real-time PCR monitoring using intrinsic imaging without labeling |
| CN102046291A (en) * | 2008-04-04 | 2011-05-04 | It-Is国际有限公司 | Thermal control system and method for chemical and biochemical reactions |
| CN102559488A (en) * | 2012-01-16 | 2012-07-11 | 福建医科大学 | Quantitative polymerase chain reaction (PCR) microfluidic chip integrated device for integrated electrochemical detection technology |
| WO2016006612A1 (en) * | 2014-07-08 | 2016-01-14 | 国立研究開発法人産業技術総合研究所 | Nucleic acid amplification device, nucleic acid amplification method, and chip for nucleic acid amplification |
| WO2017199933A1 (en) * | 2016-05-18 | 2017-11-23 | 日本板硝子株式会社 | Reaction treatment device, and method for controlling reaction treatment device |
| CN106052198A (en) * | 2016-05-31 | 2016-10-26 | 大连海事大学 | PCR amplifier with fast temperature raising and lowering function |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113201454A (en) * | 2021-05-25 | 2021-08-03 | 江南大学 | Small-size nucleic acid rapid annealing device using liquid exchange method |
| CN114608353A (en) * | 2022-03-04 | 2022-06-10 | 通用生物(滁州)有限公司 | Temperature compensation system of oligonucleotide production reaction system |
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Application publication date: 20190705 |