CN107460108A - A kind of PCR reaction systems of continuous sample introduction are prepared and sampling device and PCR instrument - Google Patents
A kind of PCR reaction systems of continuous sample introduction are prepared and sampling device and PCR instrument Download PDFInfo
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- CN107460108A CN107460108A CN201710603515.4A CN201710603515A CN107460108A CN 107460108 A CN107460108 A CN 107460108A CN 201710603515 A CN201710603515 A CN 201710603515A CN 107460108 A CN107460108 A CN 107460108A
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- 238000006243 chemical reaction Methods 0.000 title claims abstract description 97
- 238000005070 sampling Methods 0.000 title claims abstract description 26
- 238000002156 mixing Methods 0.000 claims abstract description 53
- 239000000203 mixture Substances 0.000 claims abstract description 34
- 239000007788 liquid Substances 0.000 claims abstract description 33
- 239000002699 waste material Substances 0.000 claims abstract description 31
- 230000001105 regulatory effect Effects 0.000 claims abstract description 28
- 238000005406 washing Methods 0.000 claims abstract description 19
- 239000006210 lotion Substances 0.000 claims abstract description 17
- 239000000872 buffer Substances 0.000 claims description 20
- 239000012530 fluid Substances 0.000 claims description 20
- 239000003570 air Substances 0.000 claims description 3
- 239000011261 inert gas Substances 0.000 claims description 3
- 239000002480 mineral oil Substances 0.000 claims description 3
- 235000010446 mineral oil Nutrition 0.000 claims description 3
- 239000011535 reaction buffer Substances 0.000 claims description 3
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 10
- 230000007850 degeneration Effects 0.000 description 7
- 238000004925 denaturation Methods 0.000 description 7
- 230000036425 denaturation Effects 0.000 description 7
- 239000000463 material Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 238000000137 annealing Methods 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 230000004087 circulation Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000012212 insulator Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 150000002972 pentoses Chemical class 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000011218 segmentation Effects 0.000 description 1
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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
- B01L7/525—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 with physical movement of samples between temperature zones
- B01L7/5255—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 with physical movement of samples between temperature zones by moving sample containers
Abstract
The invention provides a kind of preparation of the PCR reaction systems of continuous sample introduction and sampling device, it includes fixed component offer portion, variable composition provider, mixing chamber, sample feeding pipe and washing lotion provider, the outlet in the fixed component offer portion connects with the mixing chamber, the outlet of the variable composition provider connects with the entrance of the mixing chamber, the outlet of the washing lotion provider connects with the entrance of the mixing chamber, the outlet of the mixing chamber is connected with sample feeding pipe, the reaction system exit of the mixing chamber is provided with reaction system outlet valve, the mixing chamber is additionally provided with waste liquid outlet, the waste liquid outlet is connected with waste liquid discharge pipe, and, waste liquid dump valve is provided with the waste liquid outlet.By the device, it may be implemented in same tubular type temperature regulating device while carry out multiple PCR reactions.
Description
Technical field
The present invention relates to molecular biology equipment, more specifically it relates to a kind of PCR reaction systems of continuous sample introduction prepare and
Sampling device and PCR instrument.
Background technology
PCR (PCR) is the underlying technology of molecular biology, and its principle is, using DNA at 95 DEG C or so
High temperature double-stranded DNA is unwind, after cooling, annealed between the primer of high concentration and the DNA profiling to unwind, then by reaction system
Temperature adjustment to polymerase activity temperature, make polymerase complementary along the direction composition along phosphoric acid to pentose (5 ' -3 ')
Chain.Actual PCR instrument based on polymerase manufacture is exactly a temperature control device, can be in denaturation temperature, renaturation temperature, elongating temperature it
Between be controlled well.
Since perkin-elmercetus companies First PCR instrument comes out, tens different producers are there is now in state
Inside and outside production and selling PCR instrument.Between short several years, PCR instrument passes through the development of several generations, constantly employs new technology, and enter
One step develops towards the direction of convenient, practical, high-intelligentization and automation.
Although round pcr obtained very big development, but all PCR instruments are all to control PCR by the following method
Reaction：PCR pipe equipped with PCR reaction systems is placed in the temperature-control hole of fixed temperature control base, and is close to the interior of temperature-control hole
Wall, the temperature of PCR pipe and its internal reaction system filled is controlled by controlling the temperature of temperature control hole wall.Although this kind of device
Generalized, however it there is some it is unconquerable the shortcomings that.
It is one of main shortcoming that time-consuming.Normal PCR reactions generally comprise 30 circulations, wherein denaturation 30s, annealing
30s, extension 30s/kb.Plus denaturation 5min, last eventually to extend 10min, expanding 1kb fragment only needs one and a half hours left side
The right time.However, when being reacted in PCR instrument, it has been found that the fragment for expanding 1kb was generally required at least over 2 hours
Time.The reason is that the position of sample is fixed in PCR instrument, therefore it is necessary to the temperature for manipulating temperature control base change to
React the temperature required for the stage proceeded to.30 circulations, every time circulation have 3 alternating temperatures, that is to say, that need to carry out 90 times
The alternating temperature of left and right.This needs to consume the substantial amounts of time close to the heating and cooling process of 100 times.
In addition to expending the time in heating and cooling process, existing PCR also have one it is huge the shortcomings that be, be not suitable for complete
Automation mechanized operation.Because circular response is carried out in fixed position, it is necessary to prepare reaction system in PCR pipe, then will be equipped with
The PCR pipe of reaction system is transferred in the reacting hole of PCR instrument.Due to being frequently necessary to prepare multiple reaction systems (for example, template is not
With, primer difference etc.), no matter system is prepared and the transfer of PCR pipe, is required for spending a large amount of manpowers, can not be accomplished full-automatic.Mesh
Before untill, also without any instrument can accomplish from PCR system be formulated into PCR reaction complete full-automation.
The content of the invention
Applicants have invented a kind of tubular type temperature regulating device, reaction system is entered from reaction tube inlet, by pipe shaft, reaction
The temperature of system and identical with the pipe shaft temperature of present position, and is reacted at such a temperature by heat transfer.
Based on the device, the invention provides a kind of preparation of the PCR reaction systems of continuous sample introduction and sampling device, it includes
Fixed component offer portion, variable composition provider, mixing chamber, sample feeding pipe and washing lotion provider, the fixed component offer portion
Outlet connects with the mixing chamber, and the outlet of the variable composition provider connects with the entrance of the mixing chamber, described to wash
The outlet of liquid provider is connected with the entrance of the mixing chamber, and the outlet of the mixing chamber is connected with sample feeding pipe, the mixing chamber
Reaction system exit be provided with reaction system outlet valve, the mixing chamber is additionally provided with waste liquid outlet, the waste liquid outlet
It is connected with waste liquid discharge pipe, also, waste liquid dump valve is provided with the waste liquid outlet.Washing lotion can be the buffering of water or polymerase
Liquid.
Prepared by using the PCR reaction systems and sampling device, user can be not subjected to prepare the heavy of PCR system
Work, especially when up to tens, sample is even hundreds of.Only need setting system to prepare parameter, by control system, refer to
Show that the dosing pump set in fixed component offer portion and variable composition provider adds the respective components of set amount into mixing chamber,
It can be achieved to be automatically finished PCR system preparation, the PCR reaction systems for preparing to obtain are exported by reacted system, sample feeding pipe
It is pumped directly in tubular type temperature regulating device, you can enter performing PCR reaction, need not artificially be participated between this process.
After a reaction system prepares and enters tubular type temperature regulating device by sample feeding pipe, washing lotion provider passes through metering
Pump fills it up with washing lotion into mixing chamber, and mixing chamber is washed, and washing lotion is discharged by waste liquid outlet and waste liquid discharge pipe, after washing
Mixing chamber continue on for the preparations of next PCR reaction systems, and obtained PCR reaction systems will be prepared and pass through reacted body
System outlet, sample feeding pipe are pumped directly in tubular type temperature regulating device.So that carried out simultaneously in same tubular type temperature regulating device
Multiple PCR reactions.
In general, in being reacted with a collection of PCR, the species and dosage of enzyme, dNTP, buffer solution etc. are all fixed, primers
It is likely to be fixed with template, it is also possible to be variable, all fixed component of species and dosage is referred to as fixed group by we
Point, the component that species or dosage can change is called variable composition.If for example, to detect some gene in different samples
Presence or absence, then enzyme, dNTP, buffer solution and primer pair are all fixed, these referred to as fixed components, and template is variable
, referred to as variable composition.
In one embodiment, the fixed component offer portion is the device equipped with the fixed component of mixing.
Preferably, the every other component in addition to variable composition is provided in the fixed component offer portion in PCR reaction systems
Mixture.
In another embodiment, the fixed component offer portion includes multiple fixed component provider and a premix
Room, the entrance exported with the premixer of each fixed component provider connect, the outlet of the premixer and institute
State the entrance connection of mixing chamber.Fixed component provider is respectively provided with each fixed component, and passes through measuring pump according to the value of setting
It is input in premixer, so as to obtain the mixture available for multiple PCR reacting doses, measuring pump in premixer is by a PCR
In the mixture input mixing chamber of the amount of reaction, the variable composition of the amount of setting is added to by the measuring pump of variable composition provider
In mixing chamber, so as to be configured to PCR reaction systems.
Specifically, the multiple fixed component provider supplies PCR reaction buffers, nucleosides to the premixer respectively
Acid, polymerase, the one kind also supplied to the premixer in template and primer pair.
In one embodiment, the multiple variable composition provider provides different primers to the mixing chamber respectively
Right or different template, or the same template of various concentrations.
In one embodiment, the device is additionally provided with buffer fluid provider, is filled in the buffer fluid provider
There is buffer fluid, the outlet of the buffer fluid provider connects with the sample feeding pipe.When a reaction system enters tubular type control
After warm device, buffer fluid provider injects a certain amount of buffer fluid by measuring pump into sample feeding pipe, with space between adjacent two
Individual reaction system, and can maintenance reaction system both ends pressure.
In one embodiment, the buffer fluid is air, inert gas or mineral oil.
The invention further relates to a kind of full-automatic PCR instrument, and it includes above-mentioned PCR reaction systems preparation and sampling device and pipe
Formula temperature regulating device, the PCR reaction systems are prepared and sampling device is connected with the tubular type temperature regulating device by the sample feeding pipe.
Brief description of the drawings
Fig. 1 is reaction system preparation and the schematic diagram of sampling device of embodiment 1；
Fig. 2 provides the schematic diagram of an example in portion for fixed component；
Fig. 3 is the schematic diagram of an example of temperature regulating device；
Fig. 4 is the viewgraph of cross-section of one example of temperature regulating device；
Fig. 5 is the viewgraph of cross-section of an example of temperature regulating device；
Fig. 6 is the schematic diagram of an example of temperature regulating device；
In accompanying drawing, the list of parts representated by each label is as follows：
1st, fixed component offer portion, 101, fixed component provider, 102, premixer, 2, variable composition provider, 3, mixed
Close room, 4, sample feeding pipe, 5, temperature control post, 6, reaction tube pipe shaft, 7, entrance, 8, outlet, 9, reaction product discharge pipe, 10, washing lotion carries
For device, 11, reaction system outlet, 12, waste liquid outlet, 13, waste liquid discharge pipe, 14, buffer fluid provider, 501, degeneration area,
502nd, annealed zone, 503, extension area, 601, denaturation section, 602, annealing section, 603, extension of section, 604, insulation short tube,
701st, outlet valve, 801, valve.
Embodiment
The principle and feature of the present invention are described below in conjunction with example, the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the present invention.
Embodiment 1
As shown in figure 1, the PCR reaction systems of the continuous sample introduction of the embodiment are prepared and sampling device includes fixed component and carried
For portion 1, variable composition provider 2, mixing chamber 3, sample feeding pipe 4 and washing lotion provider 10, the outlet in the fixed component offer portion 1
Connected with the mixing chamber 3, the outlet of the variable composition provider 2 connects with the entrance of the mixing chamber 3, the washing lotion
The outlet of provider 10 is connected with the entrance of the mixing chamber 3, and the outlet 11 of the mixing chamber 3 is connected with sample feeding pipe 4, described mixed
Close and be provided with reaction system outlet valve at the reaction system outlet 11 of room 3, the mixing chamber 3 is additionally provided with waste liquid outlet 12, institute
State waste liquid outlet 12 to be connected with waste liquid discharge pipe 13, also, waste liquid dump valve is provided with the waste liquid outlet 12.The fixation
Component offer portion 1 is the device equipped with the fixed component of mixing.
Preferably, the every other component in addition to variable composition is provided in the fixed component offer portion 1 in PCR reaction systems
Mixture.
Prepared by using the PCR reaction systems and sampling device, user can be not subjected to prepare the heavy of PCR system
Work, especially when up to tens, sample is even hundreds of.Only need setting system to prepare parameter, by control system, refer to
Show that the dosing pump set in fixed component offer portion and variable composition provider adds the respective components of set amount into mixing chamber,
It can be achieved to be automatically finished PCR system preparation, the PCR reaction systems for preparing to obtain are exported by reacted system, sample feeding pipe
It is pumped directly in tubular type temperature regulating device, you can enter performing PCR reaction, need not artificially be participated between this process.
After a reaction system prepares and enters tubular type temperature regulating device by sample feeding pipe, washing lotion provider passes through metering
Pump fills it up with washing lotion into mixing chamber, and mixing chamber is washed, and washing lotion is discharged by waste liquid outlet and waste liquid discharge pipe, after washing
Mixing chamber continue on for the preparations of next PCR reaction systems, and obtained PCR reaction systems will be prepared and pass through reacted body
System outlet, sample feeding pipe are pumped directly in tubular type temperature regulating device.So that carried out simultaneously in same tubular type temperature regulating device
Multiple PCR reactions.
In general, in being reacted with a collection of PCR, the species and dosage of enzyme, dNTP, buffer solution etc. are all fixed, primers
It is likely to be fixed with template, it is also possible to be variable, all fixed component of species and dosage is referred to as fixed group by we
Point, the component that species or dosage can change is called variable composition.If for example, to detect some gene in different samples
Presence or absence, then enzyme, dNTP, buffer solution and primer pair are all fixed, these referred to as fixed components, and template is variable
, referred to as variable composition.
Embodiment 2
As illustrated in fig. 1 and 2, the reaction system of the present embodiment is prepared and sampling device includes fixed component offer portion 1, variable
Component provider 2, mixing chamber 3, sample feeding pipe 4 and washing lotion provider 10, the outlet in the fixed component offer portion 1 mix with described
Room 3 connects, and the outlet of the variable composition provider 2 connects with the entrance of the mixing chamber 3, the washing lotion provider 10
Outlet connected with the entrance of the mixing chamber 3, the outlet 11 of the mixing chamber 3 is connected with sample feeding pipe 4, the mixing chamber 3 it is anti-
Answer and be provided with reaction system outlet valve at system outlet 11, the mixing chamber 3 is additionally provided with waste liquid outlet 12, the waste liquid outlet
12 are connected with waste liquid discharge pipe 13, also, are provided with waste liquid dump valve at the waste liquid outlet 12.
The fixed component offer portion 1 includes multiple fixed component provider 101 and a premixer 102, each described
Entrance of the outlet of fixed component provider 101 with the premixer 102 connects, the outlet of the premixer 102 with it is described
The entrance connection of mixing chamber 3.Fixed component provider is respectively provided with each fixed component, and defeated by measuring pump according to the value of setting
Enter into premixer, so as to obtain the mixture available for multiple PCR reacting doses, the measuring pump in premixer is anti-by a PCR
In the mixture input mixing chamber for the amount answered, the variable composition of the amount of setting is added to mixed by the measuring pump of variable composition provider
Close in room, so as to be configured to PCR reaction systems.
Specifically, the multiple fixed component provider 101 respectively to the premixer 102 supply PCR reaction buffers,
Nucleotides, polymerase, the one kind also supplied to the premixer 102 in template and primer pair.
In one embodiment, the multiple variable composition provider 2 provides different draw to the mixing chamber 3 respectively
Thing pair, or different templates, or the same template of various concentrations.
Embodiment 3
As shown in figure 1, the device of the present embodiment is additionally provided with buffer fluid provider 14, the buffer fluid provider 14
In buffer fluid is housed, the outlet of the buffer fluid provider 14 connects with the sample feeding pipe 4.When a reaction system enters
After tubular type temperature regulating device, buffer fluid provider injects a certain amount of buffer fluid by measuring pump into sample feeding pipe, to separate
Two neighboring reaction system, and can maintenance reaction system both ends pressure.
In one embodiment, the buffer fluid is air, inert gas or mineral oil.
The invention further relates to a kind of full-automatic PCR instrument, and it includes above-mentioned PCR reaction systems preparation and sampling device and pipe
Formula temperature regulating device, the PCR reaction systems are prepared and sampling device is connected with the tubular type temperature regulating device by the sample feeding pipe 4
It is logical.
Embodiment 3
The full-automatic PCR instrument of the present invention includes above-mentioned PCR reaction systems preparation and sampling device and tubular type temperature control dress
Put, the PCR reaction systems are prepared and sampling device is connected with the tubular type temperature regulating device by the sample feeding pipe 4.
As shown in figure 3, the temperature regulating device includes temperature control post 5 and reaction tube, the reaction tube includes entrance 7, outlet 8 and pipe
Body 6, the pipe shaft 6 of the reaction tube are spirally coiled in 5 on the temperature control post, and the entrance 7 connects with sample feeding pipe 4, the outlet 8
Connected with reaction product discharge pipe 9.
In running, reaction system is entered by sample feeding pipe 4 from entrance 7, and by pipe shaft 6, the temperature of reaction system is led to
Cross heat transfer and identical with the temperature of pipe shaft 6 of present position, and reacted at such a temperature, completed after reaction from reaction product
Discharge pipe 9 comes out.Such tubular type temperature regulating device can be used for the reaction environment for building isothermal reaction and temperature cycles reacting condition.
In another embodiment, as shown in figure 4, the periphery of the cross section of temperature control post 5 of temperature regulating device is divided into three
Heated at constant temperature area, degeneration area 501, annealed zone 502 and extension area 503 being followed successively by, the temperature of the degeneration area 501 is 90-98 DEG C,
The temperature of the annealed zone 502 is 45-70 DEG C, and the temperature of the extension area 503 is 70-75 DEG C, the entrance of the reaction tube 7
In one end by the annealed zone 503 of the degeneration area 501, the outlet 8 of the reaction tube is located at the extension area 503 and leans on institute
State one end of degeneration area 501.The device can be used in PCR instrument, the temperature regulating device as PCR reactions.
The division in three heated at constant temperature areas to be formed the temperature needed for the three phases of a PCR cycle in reaction tube
Degree, and in the temperature regulating device, these three warm areas are all constant temperature, it is not necessary to denaturation temperature-annealing temperature-elongating temperature it
Between carry out cyclic switching.Reaction system flows successively through annealed zone, extension area since degeneration area, and the temperature for completing a PCR is followed
Ring, so as to often once complete the PCR reactions of a circulation around the operation of temperature control post.
In another embodiment, heated at constant temperature area may include multiple heated at constant temperature units, part heated at constant temperature unit
Degeneration area is formed, part forms annealed zone, and part forms extension area.Can be by changing the quantity of the heated at constant temperature unit in each area
To adjust the length in each area, so as to control the reaction time.
In another embodiment, temperature control post and pipe shaft can be divided into three sections, and one section is used for the initial denaturation stage, one section with
In the circular response stage, also one section is used for the last extension stage eventually.
In another embodiment, as shown in figure 5, the pipe shaft 6 of temperature regulating device reaction tube is according to residing heated at constant temperature area
Segmentation, often enclose spiral and correspondingly form three constant temperature sections, be followed successively by denaturation section 601, annealing section 602 and extension of section
603, and connected between adjacent section by short tube made of heat insulator 604.By being adopted between adjacent section
Take the short tube 604 of heat insulation to connect, preferably avoid the heat transfer between the different constant temperature sections of pipe shaft 6.In the present embodiment
So-called heat insulator refers to the relatively low material of pyroconductivity, rather than definitely athermanous material.
In a preferred scheme, the surface of temperature control post 5 has the bung flange to match with the reaction tube pipe shaft 6
Groove, the reaction tube pipe shaft 6 are embedded in the bung flange groove.The setting makes it possible to more preferably precisely control reaction tube pipe
The temperature of body 6.It is highly preferred that separated between heated at constant temperature area corresponding to the bung flange groove by heat insulator, preferably every
From the heat transfer between adjacent heated at constant temperature area.So-called heat insulator refers to the relatively low material of pyroconductivity in the present embodiment
Material, rather than definitely athermanous material.
In another embodiment, as shown in fig. 6, the pipe shaft has multiple outlets 8, it is located at respectively away from the entrance
At 7 different bung flange numbers, and each outlet 8 is both provided with outlet valve 801, and the outlet 8 farthest from entrance 7 can be not provided with exporting
Valve 801.The outlet valve 801 causes residing exit to have two states, and a state is reaction system to next spiral shell
Circle movement, another state is the pipe shaft 6 that reaction system leaves reaction tube, is come out from outlet.For example, reaction tube pipe shaft 6 can be
30-60 circles are coiled on temperature control post 1, and outlet 8 is set at the number of turns different away from entrance, setting the state of outlet valve 801 makes instead
System is answered to be come out by the specific number of turns and from corresponding outlet 8 by the reaction product discharge pipe 9 with the outlet, so as to
Carry out specific PCR reaction cycle numbers.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc., it should be included in the scope of the protection.
Claims (9)
1. a kind of PCR reaction systems of continuous sample introduction are prepared and sampling device, it is characterised in that including fixed component offer portion
(1), variable composition provider (2), mixing chamber (3), sample feeding pipe (4) and washing lotion provider (10), the fixed component offer portion
(1) outlet connects with the mixing chamber (3), the outlet of the variable composition provider (2) and entering for the mixing chamber (3)
Mouth connection, the outlet of the washing lotion provider (10) connect with the entrance of the mixing chamber (3), the outlet of the mixing chamber (3)
(11) it is connected with sample feeding pipe (4), reaction system outlet (11) place of the mixing chamber (3) is provided with reaction system outlet valve, institute
State mixing chamber (3) and be additionally provided with waste liquid outlet (12), the waste liquid outlet (12) is connected with waste liquid discharge pipe (13), also, institute
State waste liquid outlet (12) place and be provided with waste liquid dump valve.
2. PCR reaction systems according to claim 1 are prepared and sampling device, it is characterised in that the fixed component carries
It is the device equipped with the fixed component of mixing for portion (1).
3. PCR reaction systems according to claim 2 are prepared and sampling device, it is characterised in that the fixed component carries
Mixture for being equipped with the every other component in addition to variable composition in PCR reaction systems in portion (1).
4. PCR reaction systems according to claim 1 are prepared and sampling device, it is characterised in that the fixed component carries
Include multiple fixed component provider (101) and a premixer (102) for portion (1), each fixed component provider
(101) entrance of the outlet with the premixer (102) connects, outlet and the mixing chamber (3) of the premixer (102)
Entrance connection.
5. PCR reaction systems according to claim 4 are prepared and sampling device, it is characterised in that the multiple fixed group
Provider (101) is divided to supply PCR reaction buffers, nucleotides, polymerase to the premixer (102) respectively, also to described pre-
One kind in mixed room (102) supply template and primer pair.
6. PCR reaction systems according to claim 1 are prepared and sampling device, it is characterised in that the multiple changeable set
Point provider (2) provides different primer pairs, or different templates to the mixing chamber (3) respectively, or various concentrations is identical
Template.
7. the PCR reaction systems according to any one of claim 1-6 are prepared and sampling device, it is characterised in that also set
Buffer fluid provider (14) is equipped with, buffer fluid is housed in the buffer fluid provider (14), the buffer fluid provides
The outlet of device (14) connects with the sample feeding pipe (4).
8. PCR reaction systems according to claim 7 are prepared and sampling device, it is characterised in that the buffer fluid is
Air, inert gas or mineral oil.
9. a kind of full-automatic PCR instrument, it is characterised in that prepared including the PCR reaction systems any one of claim 1-8
And sampling device and tubular type temperature regulating device, the PCR reaction systems are prepared and sampling device leads to the tubular type temperature regulating device
Cross the sample feeding pipe (4) connection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710603515.4A CN107460108A (en) | 2017-07-23 | 2017-07-23 | A kind of PCR reaction systems of continuous sample introduction are prepared and sampling device and PCR instrument |
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| CN201710603515.4A CN107460108A (en) | 2017-07-23 | 2017-07-23 | A kind of PCR reaction systems of continuous sample introduction are prepared and sampling device and PCR instrument |
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Citations (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1377639A (en) * | 1972-12-13 | 1974-12-18 | Vitafoam Ltd | Compounding and dispensing apparatus |
| EP0039856A1 (en) * | 1980-05-14 | 1981-11-18 | Bayer Ag | Apparatus for producing a reactive mixture to form a homogeneous material or foam from at least two inter-reacting components |
| JPH10304890A (en) * | 1997-05-02 | 1998-11-17 | Biomerieux Vitek Inc | Improved nucleic acid assay |
| AU5706999A (en) * | 1997-05-02 | 1999-12-23 | Biomerieux Vitek, Inc. | Dual chamber disposable reaction vessel for amplification reactions, reaction processing station therefor, and methods of use |
| JP2007101200A (en) * | 2005-09-30 | 2007-04-19 | Yokogawa Electric Corp | Cartridge for chemical reaction and its usage |
| CN101184983A (en) * | 2005-03-16 | 2008-05-21 | 雅拓晶科生物系统（私人）有限公司 | Methods and device for transmitting, enclosing and analysing fluid samples |
| JP2008278810A (en) * | 2007-05-11 | 2008-11-20 | Sony Corp | Real-time pcr apparatus |
| CN102304464A (en) * | 2011-10-08 | 2012-01-04 | 武汉馨世生物科技有限公司 | Multiplex polymerase chain reaction amplification instrument |
| CN102911855A (en) * | 2012-10-19 | 2013-02-06 | 天津金思德生物技术有限公司 | Warm start method of PCR (Polymerase Chain Reaction) instrument and PCR instrument with warm start function |
| CN202881280U (en) * | 2012-11-15 | 2013-04-17 | 王波 | Real-time fluorescent quantitative PCR (polymerase chain reaction) instrument |
| CN103602659A (en) * | 2013-10-23 | 2014-02-26 | 杨锦宇 | High-throughput nucleic acid continual amplification apparatus and method |
| CN103667046A (en) * | 2013-12-19 | 2014-03-26 | 江苏金太生命科技有限公司 | PCR (polymerase chain reaction) instrument with partitions |
| CN105039155A (en) * | 2015-07-29 | 2015-11-11 | 杭州安杰思生物科技有限公司 | Real-time fluorescent quantitative PCR instrument |
| CN105349401A (en) * | 2015-10-14 | 2016-02-24 | 安徽易康达光电科技有限公司 | Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof |
| CN108048315A (en) * | 2018-01-04 | 2018-05-18 | 深圳大学 | It is a kind of based on the fluorescence quantitative PCR instrument being automatically loaded |
| US20190062802A1 (en) * | 2017-08-28 | 2019-02-28 | United States Of America As Represented By The Administrator Of Nasa | Rapid nucleic isolation method and fluid handling devices |
| CN112029653A (en) * | 2020-08-17 | 2020-12-04 | 浙江大学 | Digital nucleic acid amplification detection method and integrated detection system based on CRISPR and Cas |
-
2017
- 2017-07-23 CN CN201710603515.4A patent/CN107460108A/en active Pending
Patent Citations (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1377639A (en) * | 1972-12-13 | 1974-12-18 | Vitafoam Ltd | Compounding and dispensing apparatus |
| EP0039856A1 (en) * | 1980-05-14 | 1981-11-18 | Bayer Ag | Apparatus for producing a reactive mixture to form a homogeneous material or foam from at least two inter-reacting components |
| JPH10304890A (en) * | 1997-05-02 | 1998-11-17 | Biomerieux Vitek Inc | Improved nucleic acid assay |
| AU5706999A (en) * | 1997-05-02 | 1999-12-23 | Biomerieux Vitek, Inc. | Dual chamber disposable reaction vessel for amplification reactions, reaction processing station therefor, and methods of use |
| CN101184983A (en) * | 2005-03-16 | 2008-05-21 | 雅拓晶科生物系统（私人）有限公司 | Methods and device for transmitting, enclosing and analysing fluid samples |
| JP2007101200A (en) * | 2005-09-30 | 2007-04-19 | Yokogawa Electric Corp | Cartridge for chemical reaction and its usage |
| JP2008278810A (en) * | 2007-05-11 | 2008-11-20 | Sony Corp | Real-time pcr apparatus |
| CN102304464A (en) * | 2011-10-08 | 2012-01-04 | 武汉馨世生物科技有限公司 | Multiplex polymerase chain reaction amplification instrument |
| CN102911855A (en) * | 2012-10-19 | 2013-02-06 | 天津金思德生物技术有限公司 | Warm start method of PCR (Polymerase Chain Reaction) instrument and PCR instrument with warm start function |
| CN202881280U (en) * | 2012-11-15 | 2013-04-17 | 王波 | Real-time fluorescent quantitative PCR (polymerase chain reaction) instrument |
| CN103602659A (en) * | 2013-10-23 | 2014-02-26 | 杨锦宇 | High-throughput nucleic acid continual amplification apparatus and method |
| CN103667046A (en) * | 2013-12-19 | 2014-03-26 | 江苏金太生命科技有限公司 | PCR (polymerase chain reaction) instrument with partitions |
| CN105039155A (en) * | 2015-07-29 | 2015-11-11 | 杭州安杰思生物科技有限公司 | Real-time fluorescent quantitative PCR instrument |
| CN105349401A (en) * | 2015-10-14 | 2016-02-24 | 安徽易康达光电科技有限公司 | Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof |
| US20190062802A1 (en) * | 2017-08-28 | 2019-02-28 | United States Of America As Represented By The Administrator Of Nasa | Rapid nucleic isolation method and fluid handling devices |
| CN108048315A (en) * | 2018-01-04 | 2018-05-18 | 深圳大学 | It is a kind of based on the fluorescence quantitative PCR instrument being automatically loaded |
| CN112029653A (en) * | 2020-08-17 | 2020-12-04 | 浙江大学 | Digital nucleic acid amplification detection method and integrated detection system based on CRISPR and Cas |
Non-Patent Citations (1)
| Title |
|---|
| 侯新东主编: "《生物化学与分子生物学实验教程》", 31 December 2016, 中国地质大学出版社 * |
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