CN106754341A - A kind of droplet type digital pcr biochip - Google Patents
A kind of droplet type digital pcr biochip Download PDFInfo
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- CN106754341A CN106754341A CN201611263102.8A CN201611263102A CN106754341A CN 106754341 A CN106754341 A CN 106754341A CN 201611263102 A CN201611263102 A CN 201611263102A CN 106754341 A CN106754341 A CN 106754341A
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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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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00279—Features relating to reactor vessels
- B01J2219/00306—Reactor vessels in a multiple arrangement
- B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
- B01J2219/00315—Microtiter plates
- B01J2219/00317—Microwell devices, i.e. having large numbers of wells
Abstract
The invention discloses a kind of droplet type digital pcr biochip, the chip body formed including being fitted with bottom sheet by upper piece, sample cavity, drop storage chamber and oil-discharging cavity are provided with inside the chip body, chip body surface is provided with the sample holes and outage for being connected with sample cavity and oil-discharging cavity respectively, it is respectively equipped with multiple droplet formation ducts and oil extraction duct between the sample cavity and drop storage chamber, between drop storage chamber and oil-discharging cavity, the height in the droplet formation duct and oil extraction duct is respectively less than the height of drop storage chamber.Chip of the present invention, when water enters into more broad drop storage chamber by more narrow droplet formation duct, water accelerates when part enters drop storage chamber because surface tension effects flow, and be mutually broken with the water in droplet formation duct, form drop.The droplet size of preparation is homogeneous, and preparation speed is fast, flux is high.
Description
Technical field
The present invention relates to biochip preparing technical field, more particularly to a kind of droplet type digital pcr biochip.
Background technology
Quantitative fluorescent PCR (Fluorescence Quantitative Polymerase Chain Reaction, qPCR)
A routine techniques for key of biology field is had evolved into, the hair of life science every field has greatly been promoted
Exhibition.But, influence the factor of its amplification efficiency many in PCR amplification procedures, it is difficult to ensure that actual sample and standard sample and
Amplification efficiency between different samples is identical, and the basis-cycle threshold (Ct values) for thus causing its quantitative analysis to be relied on is not
Invariable.Therefore qPCR's is quantitatively relative quantification, and its degree of accuracy and reappearance can not still meet molecular biosciences
Learn the requirement of quantitative analysis.Further, since inhibitory action of the pcr amplification product to enzymic catalytic reaction, is currently based on qPCR technologies
Genetic mutation detection method it is usually helpless to low-abundance genetic mutation in body cell.
Digital pcr (Digital PCR, dPCR) is a kind of nucleic acid quantification counted based on single-molecule PCR method
Method, is a kind of method of absolute quantitation.The main micro-fluidic or droplet side using present analysis chemistry hot topic research field
Method, the nucleic acid solution after Macrodilution is dispersed in the microreactor of chip or droplet, the nucleic acid-templated number of each reactor
Less than or equal to 1.After so by PCR cycle, the reactor for having nucleic acid templates will provide fluorescence signal, not have
There is the reactor of template just without fluorescence signal.According to relative scale and the volume of reactor, it is possible to extrapolate original solution
Nucleic acid concentration.It is different from traditional quantitative PCR, method of the digital pcr by directly counting, it is possible to achieve initial DNA profiling
Absolute quantitation.
In addition, digital pcr or a kind of side that micro mutant can be identified in substantial amounts of wild type DNA backgrounds
Method.Individually expanded because digital pcr technology can in advance separate template DNA molecule, avoiding problems high abundance
Amplification of the allele nucleic acid to variant nucleic acid suppresses, therefore improves the detector efficiency of micro variant nucleic acid.Emulsion droplet digital pcr
Technology can detect as little as 0.001% mutant fragments, and sequencing and conventional quantitative real-time PCR are to prominent less than 1%
Change is helpless, therefore abrupt climatic change sensitivity can be improved 1000 times by droplet type digital pcr technology.
The general operation process CIMS of traditional digital pcr technology is complicated, typically first by manual stepwise dilution simultaneously
In distribution sample to microwell plate, microwell plate is then placed in the enterprising performing PCR reaction of thermal cycler, reaction is read after terminating by instrument
The fluorescence signal in each micropore is taken, finally according to Poisson distribution principle and the ratio of positive droplet, binding analysis software can
Calculating provides the concentration or copy number of target molecule to be checked.The liquid that this mode is cumbersome, flux is small, efficiency is low and less
Drop is decomposed number and also limit its precision and measurable dynamic range, and application aspect has great limitation.In recent years, micro-fluidic skill
The development for developing into digital pcr technology of art is filled with new vitality.Because microflow control technique is excellent in terms of microfluid manipulation
Gesture so that sample can be decomposed into nanoliter even picoliters level by us, obtained more samples and decomposed numbers, so as to greatly improve
The detection sensitivity of digital pcr technology, confidence level and dynamic range degree.In addition, microflow control technique automation, easy of integration, flux
Advantage high, can also be greatly enhanced the detection efficiency of digital pcr technology.
By advantage of the microflow control technique in terms of microfluid manipulation, existing multiple research group and company in recent international
Develop the digital pcr system based on microflow control technique.Compare be typically include Fluidigm companies release based on microchamber type
BioMarkTM systems and Bio-Rad companies release the QX100TM systems based on drop type.
Microlayer model technology refers to form Water-In-Oil or oil-in-water metastable independent microbody hydrops using immiscible phase
Drop.Emulsifying manner has various, wherein the drop formation technology based on micro-fluidic chip is rapidly developed recent years, extensively should
It is a kind of very important technology for living nature and material circle.Its principle is mainly by certain angle between two-phase liquid stream
Under house type squeezing action, wherein a phase continuous flow breaks to form drop.Conventional drop preparation method has orthohormbic structure at present
(T-junction) focused on (Flow-focusing) etc. with streaming.The systematic comparison of these method integral devices is complicated, to fluid
The requirement of control system is higher.
The content of the invention
The present invention is in view of the shortcomings of the prior art, there is provided a kind of droplet type digital pcr biochip, droplet type numeral
PCR biochips can be used for a large amount of uniform drops of quick preparation, and drop is laid on chip, only need to be to core during detection
Piece integrally carries out image checking, it is not necessary to the detection of single is carried out to drop as flow cytometer.
Droplet type digital pcr biochip, including fitted with bottom sheet by upper piece chip body for being formed, the chip body
Inside is provided with sample cavity, drop storage chamber and oil-discharging cavity, and chip body surface is provided with and is connected with sample cavity and oil-discharging cavity respectively
Sample holes and outage, be respectively equipped between the sample cavity and drop storage chamber, between drop storage chamber and oil-discharging cavity many
The height in individual droplet formation duct and oil extraction duct, the droplet formation duct and oil extraction duct is respectively less than the height of drop storage chamber
Degree so that droplet formation duct has the ledge structure for forming drop with the junction of drop storage chamber.
When using, first pass through sample holes and oil phase is injected into chip internal, treat that chip internal fills oil phase, exclude air
Afterwards, water phase is injected by sample holes, under pressure, water enters droplet formation duct from sample cavity, then from drop
When formation duct out enters into drop storage chamber, because the height in droplet formation duct is small compared with drop storage chamber, in drop shape
Step is formed into duct and drop storage chamber intersection, is entered into by more narrow droplet formation duct in water more broad
Drop storage chamber when, water part enter drop storage chamber when, due to surface tension effects flow accelerate, and with drop shape
Mutually it is broken into the water in duct, forms drop, the method is referred to as ladder emulsification method.Ladder emulsification method can be using single
One driving source, continuously generates drop, and the size of drop is mainly determined by the configuration of surface tension and micro-structural, by the shadow of flow velocity
Sound is smaller.Drop is produced after entering drop storage chamber, and the oil phase of respective volume is just discharged to oil-discharging cavity by oil extraction duct, final warp
The outage discharge connected with oil-discharging cavity.Oil extraction duct is sufficiently small so that drop can not be discharged from oil extraction duct.Produced
A large amount of drops are finally laid in drop storage chamber.After the completion of prepared by drop, the chip directly can be placed into corresponding PCR anti-
Answer and enter in instrument performing PCR to expand, after the completion of amplification, directly the chip is positioned in chip analysis instrument carries out fluorescence signal
Imaging and reading.
Preferably, the drop storage chamber is U-shape structure, and the sample cavity is linear structure, located at the two of U-shape structure
Between arm, and the droplet formation duct of multiple parallel arrangements is provided between two-arm；The oil-discharging cavity is located at the U-shape structure
Periphery.So design causes that the sample cavity of strip is stretched into inside drop storage chamber, in the top of sample cavity and both sides long
A large amount of droplet formation ducts can be set, flux prepared by drop is improved, meanwhile, also cause each border of drop storage chamber not
Can from droplet formation duct too away from, it is to avoid the drop of generation migration distance too far and destroy and lose.
It is further preferred that the sample holes are sample introduction end near the end of U-shape structure opening, sample introduction end and sample holes it
Between be provided with long and narrow joining section.
Still more preferably, the arrangement density in the droplet formation duct gradually increases from sample introduction end initial density
Greatly.This arrangement for forming certain gradient, can approach the speed for producing drop everywhere.This design can avoid passage from uniformly dividing
The channel pressure close to sample introduction end caused by cloth is excessive, and aqueous phase flow rate is too fast, produces drop excessive.
Preferably, the end of the droplet formation duct connection drop storage chamber is provided with circular arc chamfering.Chamfering is set herein
Also for generation and movement beneficial to drop.
Still more preferably, length of the circular arc chamfering on droplet formation direction is 1~100 μm.The circular arc
Length scale of the chamfering on droplet formation direction can influence generate drop size, within the specific limits, droplet size with
This length increases and increases.
Preferably, the cross section in the droplet formation duct is rectangle, and width is 10~200 μm, is highly 1~100 μm,
Drop storage chamber is highly 10~200 μm.The size of drop has direct relation, droplet formation hole with the size in droplet formation duct
The width in road is bigger with the drop of the bigger generation of height, so the size in droplet formation duct is needed in suitable scope.
It is further preferred that the droplet formation passage becomes big near drop storage chamber one end width forms bell mouth shape.
Bell mouth shape is conducive to the generation and movement of drop.
Preferably, the sample holes entrance is provided with the sealed interface plug of silica gel material, and the sealed interface plug is communicated with
The through hole of sample cavity.Silica gel material has good elasticity, using have resilient sealed interface plug can ensure to refuel mutually and
During water phase, pipette can be fitted close with sealing-plug, it is ensured that seal during sample-adding in chip runner, make flow velocity and pressure all
Energy steady implementation, promotes the uniformity of droplet formation.
Preferably, described chip body is provided with multiple reaction zones, and sample cavity, drop are equipped with inside each reaction zone
Storage chamber and oil-discharging cavity, chip body surface are provided with the sample holes and outage for being connected with sample cavity and oil-discharging cavity respectively, institute
State between sample cavity and drop storage chamber, multiple droplet formation ducts and oil extraction are respectively equipped between drop storage chamber and oil-discharging cavity
The height in duct, the droplet formation duct and oil extraction duct is respectively less than the height of drop storage chamber.It is integrated on one chip
Multiple reaction zones can increase the flux of reaction, conveniently carry out the experiment of multiple samples.
Droplet type digital pcr biochip of the present invention is small compared with drop storage chamber by the height in droplet formation duct, so that
Step is formed therebetween, and more broad drop storage chamber is entered into by more narrow droplet formation duct in water
When, water part enter drop storage chamber when, due to surface tension effects flow accelerate, and with droplet formation duct in water
Mutually it is broken, forms drop.The drop for being generated is capable of the drop storage chamber for being laid in chip of stable and uniform, without to liquid
Drop is shifted just directly can be tested and be tested and analyzed on chip to drop, greatly reduce operating procedure, be simplified
Operating system；And prepared droplet size is homogeneous, the speed for preparing drop is fast, flux is high, substantially reduces drop preparation
Time.
Brief description of the drawings
Fig. 1 is the cross-sectional view of droplet type digital pcr biochip of the present invention；
Fig. 2 be upper piece be located at chip internal one side structural representation；
Fig. 3 is cross-sectional view of the droplet type digital pcr biochip of the present invention by A-A directions in Fig. 1；
Fig. 4 is B partial enlarged drawings in Fig. 2；
Fig. 5 is the partial structural diagram in droplet formation duct and drop storage chamber；
Fig. 6 is drop figure in embodiment 3；
Fig. 7 is drop fluorescence signal detection figure in embodiment 3.
Specific embodiment
Embodiment 1
As shown in Fig. 1~5, a kind of droplet type digital pcr biochip, including by upper piece 1 and bottom sheet 2 fit the core that is formed
Piece body, is provided with sample cavity 3, drop storage chamber 4 and oil-discharging cavity 5 on upper 1 inside the chip body.
Drop storage chamber 4 is U-shape structure, and sample cavity 3 is linear structure, between U-shape structure two-arm, and and two-arm
Between be provided with the droplet formation ducts 6 of multiple parallel arrangements；Oil-discharging cavity 5 is located at the periphery of the U-shape structure.Drop storage chamber 4
Multiple oil extraction ducts 7 are provided between oil-discharging cavity 5.The height in droplet formation duct 6 and oil extraction duct 7 is respectively less than drop storage chamber
4 height so that droplet formation duct 6 has the ledge structure for forming drop with the junction of drop storage chamber 4.
Drop storage chamber 4 causes that the sample cavity 3 of strip is stretched into inside drop storage chamber 4 for U-shape structure, in sample cavity 3
Top and both sides long long a large amount of droplet formation ducts 6 can be set, increase speed prepared by drop, meanwhile, also cause liquid
Drip storage chamber 4 each border will not from droplet formation duct 6 too away from, it is to avoid the drop of generation migration distance too far and occur
Destruction and loss.
Chip body surface is provided with the sample holes 8 and outage 9 for being connected with sample cavity 3 and oil-discharging cavity 5 respectively.Sample holes 8
End near U-shape structure opening is sample introduction end, and long and narrow joining section 10 is provided between sample introduction end and sample holes 8.Droplet formation
The arrangement density in duct 6 gradually increases from sample introduction end initial density.This arrangement for forming certain gradient, can make to produce everywhere
The speed of drop is approached.If if uniform arrangement, droplet formation duct 6 close to sample introduction end is larger due to pressure, water
Phase flow velocity is very fast, and generation drop can be more.
The end that droplet formation duct 6 connects drop storage chamber 4 is provided with circular arc chamfering 11.Circular arc chamfering 11 is in droplet formation
Length on direction is 1~100 μm.Length scale of the circular arc chamfering 11 on droplet formation direction can influence to generate the big of drop
Small, within the specific limits, droplet size increases as this length increases.
The cross section in droplet formation duct 6 is rectangle, and width is 10~200 μm, is highly 1~100 μm, drop storage chamber
Highly it is 10~200 μm.Droplet formation duct 6 becomes big and forms bell mouth shape near one end width of drop storage chamber 4.Drop it is big
Small to have direct relation with droplet formation duct 6 size, the drop of the bigger generation in droplet formation duct 6 is bigger, so drop
The size for forming duct 6 is needed in suitable scope.Bell mouth shape is conducive to the generation and movement of drop.
The inlet seal of sample holes 8 has the sealing-plug of silica gel material.Silica gel material has good elasticity, using with elasticity
Sealing-plug can ensure to refuel mutually and during water phase, pipette can be fitted close with sealing-plug, it is ensured that close in chip runner
Closing property, make flow velocity and pressure can steady implementation, promote the uniformity of droplet formation.
Chip of the present invention is also carved with known to a series of sizes in sample cavity 3 to be justified as scale, can be under the microscope
During observation, the droplet size prepared by convenient judgement.
When using, oil phase is first injected into chip internal by sample holes, treats that chip internal fills oil phase, exclude air
Afterwards, water phase is injected by sample holes 8, under pressure, water enters droplet formation duct 6 from sample cavity 3, then in liquid
When drop forms duct 6 and out enters into drop storage chamber 4, because the height in droplet formation duct 6 is small compared with drop storage chamber 4,
Droplet formation duct 6 forms step with the intersection of drop storage chamber 4, is entered by more narrow droplet formation duct 6 in water
During to more broad drop storage chamber 4, water adds when part enters drop storage chamber 4 because surface tension effects flow
Speed, and be mutually broken with the water in droplet formation duct 6, form drop.Drop is produced after entering drop storage chamber 4, accordingly
The oil phase of volume is just discharged to oil-discharging cavity 5 by oil extraction duct 7, and the final outage 9 through being connected with oil-discharging cavity 5 is discharged.Outage
Height of the height of road 7 less than drop storage chamber 4 so that drop is not easy to be discharged from oil extraction duct 7.Produced a large amount of drops are most
After be laid in drop storage chamber 4, drop is Water-In-Oil structure, and the oil phase of outer layer is each other Fusion Strain, equivalent to
There are many drops of separate water phase in oil phase.After the completion of prepared by drop, directly the chip can be placed into
Enter performing PCR amplification in corresponding PCR reaction kits, after the completion of amplification, directly be positioned in chip analysis instrument by the chip
The imaging and reading of row fluorescence signal.
Embodiment 2
Described chip body is provided with multiple reaction zones, and sample cavity 3, drop storage chamber are equipped with inside each reaction zone
4 and oil-discharging cavity 5, chip body surface is provided with the sample holes 8 and outage 9 for being connected with sample cavity 3 and oil-discharging cavity 5 respectively, enters
Multiple droplet formation ducts 6 and row are respectively equipped between sample chamber 3 and drop storage chamber 4, between drop storage chamber 4 and oil-discharging cavity 5
The height in oilhole road 7, droplet formation duct 6 and oil extraction duct 7 is respectively less than the height of drop storage chamber 4.Remaining structure is with implementation
Example 1.
Embodiment 3
Using the droplet type digital pcr biochip in embodiment 1, use the n-tetradecane+3% of 70% mineral oil+30%
Used as oil phase composition, (ratio of mineral oil and n-tetradecane refers to the quality that accounts for bulk composition to EM90+3%TritonX-100
Ratio, and the ratio of EM90 and TritonX-100 is the ratio of respective quality and bulk composition.) drop preparation experiment is carried out, and
Enter performing PCR amplification.Idiographic flow is as follows：
(1) oil phase is prepared.
(2) it is mutually PCR reaction systems to prepare water.
Template comes from non-small cell lung cancer (NSCLC) cell line H1975, while there is two kinds of mutation of T790M and L858R.
Primer sequence is：
F：5’-GCCTGCTGGGCATCTG-3’；
R：5’-TCTTTGTGTTCCCGGACATAGAC-3’
Probe sequence is：
5 '-FAM-ATGAGCTGCATGATGAG-MGB-NFQ-3 ', wherein FAM are fluorescent reporter group, and NFQ quenches for fluorescence
Go out group.
Water is mutually formulated：
| 2 × PCR reaction buffers (including Taq enzyme, dNTP, magnesium ion) | 7.5uL |
| BSA (1%) | 1.5uL |
| Sense primer F (10 μM) | 0.3uL |
| Anti-sense primer R (10 μM) | 0.3uL |
| Probe (5 μM) | 0.3uL |
| Template (5ng/ μ L) | 1.0uL |
| Deionized water | 4.1uL |
| Cumulative volume | 15uL |
(3) oil phase is laid in droplet type digital pcr biochip.
(4) drop is formed simultaneously in water is added into oil phase with syringe pump by the way of ladder is emulsified.
(5) generated droplet enters performing PCR and expands by following program：
96 DEG C of predegeneration 10min；98 DEG C of denaturation 30s, 62 DEG C of annealing extend 1min, totally 39 circulations；62 DEG C of extension 1min,
25 DEG C of insulations.
(6) amplification terminates, micro- sem observation droplet morphology.If drop stable homogeneous, reading apparatus detection fluorescence signal.
As a result：After PCR amplifications, droplet size is homogeneous, and liquid-drop diameter does not crush and merge drop at 80 μm or so, substantially
(Fig. 6), instrument can detect strong fluorescence signal (Fig. 7).
SEQUENCE LISTING
<110>Use and reach bio tech ltd in Hangzhou
<120>A kind of droplet type digital pcr biochip
<130>
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 16
<212> DNA
<213>Artificial sequence
<400> 1
gcctgctggg catctg 16
<210> 2
<211> 23
<212> DNA
<213>Artificial sequence
<400> 2
tctttgtgtt cccggacata gac 23
<210> 3
<211> 17
<212> DNA
<213>Artificial sequence
<400> 3
atgagctgca tgatgag 17
Claims (10)
1. droplet type digital pcr biochip, including the chip body for being formed of being fitted with bottom sheet by upper piece, it is characterised in that institute
State and be provided with inside chip body sample cavity, drop storage chamber and oil-discharging cavity, chip body surface be provided with respectively with sample cavity and
The sample holes and outage of oil-discharging cavity connection, between the sample cavity and drop storage chamber, between drop storage chamber and oil-discharging cavity
It is respectively equipped with multiple droplet formation ducts and oil extraction duct, the height in the droplet formation duct and oil extraction duct is respectively less than drop
The height of storage chamber so that droplet formation duct has the ledge structure for forming drop with the junction of drop storage chamber.
2. droplet type digital pcr biochip as claimed in claim 1, it is characterised in that the drop storage chamber is U-shaped knot
Structure, the sample cavity is linear structure, between the two-arm of U-shape structure, and multiple parallel arrangements is provided between two-arm
Droplet formation duct；The oil-discharging cavity is located at the periphery of the U-shape structure.
3. droplet type digital pcr biochip as claimed in claim 2, it is characterised in that the sample holes are near U-shape structure
The end of opening is sample introduction end, and long and narrow joining section is provided between sample introduction end and sample holes.
4. droplet type digital pcr biochip as claimed in claim 3, it is characterised in that the row in the droplet formation duct
Cloth density gradually increases from sample introduction end initial density.
5. droplet type digital pcr biochip as claimed in claim 1, it is characterised in that the droplet formation duct connection
The end of drop storage chamber is provided with circular arc chamfering.
6. droplet type digital pcr biochip as claimed in claim 5, it is characterised in that the circular arc chamfering is in drop shape
It it is 1~100 μm into the length on direction.
7. droplet type digital pcr biochip as claimed in claim 1, it is characterised in that the horizontal stroke in the droplet formation duct
Rectangular cross-section, width is 10~200 μm, is highly 1~100 μm, and drop storage chamber is highly 10~200 μm.
8. droplet type digital pcr biochip as claimed in claim 7, it is characterised in that the droplet formation passage is close to
Drop storage chamber one end width becomes big and forms bell mouth shape.
9. drop biotinylated biomolecule chip as claimed in claim 1, it is characterised in that the sample holes entrance is provided with silica gel material
Sealed interface plug, the sealed interface plug is communicated with the through hole of sample cavity.
10. drop biotinylated biomolecule chip as claimed in claim 1, it is characterised in that described chip body is provided with multiple
Reaction zone, is equipped with sample cavity, drop storage chamber and oil-discharging cavity inside each reaction zone, chip body surface be provided with respectively with
Sample holes and outage that sample cavity is connected with oil-discharging cavity, between the sample cavity and drop storage chamber, drop storage chamber with row
It is respectively equipped with multiple droplet formation ducts and oil extraction duct between oil pocket, the height in the droplet formation duct and oil extraction duct is equal
Less than the height of drop storage chamber.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611263102.8A CN106754341B (en) | 2016-12-30 | 2016-12-30 | A kind of droplet type digital pcr biochip |
| PCT/CN2017/113847 WO2018099420A1 (en) | 2016-11-30 | 2017-11-30 | Droplet digital pcr chip |
| US16/465,438 US11376595B2 (en) | 2016-11-30 | 2017-11-30 | Droplet digital PCR chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611263102.8A CN106754341B (en) | 2016-12-30 | 2016-12-30 | A kind of droplet type digital pcr biochip |
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| CN106754341A true CN106754341A (en) | 2017-05-31 |
| CN106754341B CN106754341B (en) | 2019-01-18 |
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| CN201611263102.8A Expired - Fee Related CN106754341B (en) | 2016-11-30 | 2016-12-30 | A kind of droplet type digital pcr biochip |
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| CN108004136A (en) * | 2017-12-07 | 2018-05-08 | 深圳市博瑞生物科技有限公司 | A kind of Microfluidic droplet generates chip |
| WO2018099420A1 (en) * | 2016-11-30 | 2018-06-07 | 领航基因科技（杭州）有限公司 | Droplet digital pcr chip |
| CN108273574A (en) * | 2018-01-23 | 2018-07-13 | 杭州凯基科技有限公司 | Centrifugal microlayer model particle chip |
| CN109207360A (en) * | 2018-09-06 | 2019-01-15 | 段学欣 | A kind of digital pcr chip and its application method and the reagent segmenting system based on the chip |
| CN109294866A (en) * | 2017-07-24 | 2019-02-01 | 广州康昕瑞基因健康科技有限公司 | Emulsion chip and emulsion preparing device |
| CN109294874A (en) * | 2018-10-29 | 2019-02-01 | 领航基因科技（杭州）有限公司 | Micro-fluidic chip, device containing the chip and application thereof, the method for preparing drop using the chip or device |
| CN109370876A (en) * | 2018-12-12 | 2019-02-22 | 深圳先进技术研究院 | A kind of drop number pcr chip and drop number PCR device |
| CN110068558A (en) * | 2018-01-24 | 2019-07-30 | 思纳福(北京)医疗科技有限公司 | Microlayer model container |
| WO2019144905A1 (en) * | 2018-01-24 | 2019-08-01 | 北京光阱管理咨询合伙企业（有限合伙） | Microdroplet container, method for preparing microdroplet container, microdroplet spreading method, microdroplet formation kit, temperature control device, oil phase composition for microdroplet formation and treatment method therefor |
| WO2020034482A1 (en) * | 2018-08-13 | 2020-02-20 | 上海新微技术研发中心有限公司 | Digital pcr system and digital pcr droplet formation method |
| WO2020034483A1 (en) * | 2018-08-13 | 2020-02-20 | 上海新微技术研发中心有限公司 | Digital pcr system and digital pcr droplet formation method |
| CN112840012A (en) * | 2018-10-01 | 2021-05-25 | Sm分子生物研究有限公司 | Micropipette tip for forming microdroplets |
| CN114369647A (en) * | 2021-12-31 | 2022-04-19 | 深圳麦科田生物医疗技术股份有限公司 | Water-in-oil droplet for micro-droplet digital PCR and application thereof |
| CN114717100A (en) * | 2021-07-16 | 2022-07-08 | 墨卓生物科技(浙江)有限公司 | Microfluidic chip for single cell sequencing and application |
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| CN114369647A (en) * | 2021-12-31 | 2022-04-19 | 深圳麦科田生物医疗技术股份有限公司 | Water-in-oil droplet for micro-droplet digital PCR and application thereof |
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