CN106190817B - Nucleic acid cleavage liquid separating method and device based on porous plate - Google Patents
Nucleic acid cleavage liquid separating method and device based on porous plate Download PDFInfo
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- CN106190817B CN106190817B CN201610579824.8A CN201610579824A CN106190817B CN 106190817 B CN106190817 B CN 106190817B CN 201610579824 A CN201610579824 A CN 201610579824A CN 106190817 B CN106190817 B CN 106190817B
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- porous plate
- building enclosure
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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Abstract
The invention belongs to the measurement of microorganism or method of inspection technical field, in particular to a kind of nucleic acid cleavage liquid separating method and device based on porous plate.The nucleic acid cleavage liquid separating method based on porous plate includes the following steps: that (1) takes filtering porous plate and accepts porous plate, be provided in the reacting hole of filtering porous plate filter membrane be connected to the channel of reacting hole interior or exterior space；(2) the upper building enclosure of set outside porous plate is being accepted；(3) filtering porous plate makes building enclosure form confined space in building enclosure top cover；(4) nucleic acid cleavage liquid is injected in the reacting hole of filtering porous plate；(5) negative pressure is formed in the confined space that building enclosure is formed, the nucleic acid cleavage liquid in filtering porous plate reacting hole is made to enter underlying undertaking porous plate through filter membrane.The application realizes the nucleic acid cleavage liquid being located in porous plate reacting hole under the premise of not generating excessive vibration and is automatically separated using porous plate as carrier.
Description
Technical field
The invention belongs to the measurement of microorganism or method of inspection technical field, in particular to a kind of nucleic acid based on porous plate
Crack liquid separating method and device.
Background technique
Before carrying out microbial molecules detection to specific sample, need sample preprocessing.By taking solid-state food as an example, at sample
The purpose of reason is by microbial cell cracking and microorganism enrichment.The nucleic acid cleavage liquid obtained after microbial cell cracking, needs
It is isolated using solid-liquid separation means.In the prior art, it is generally separated using centrifuge.As the People's Republic of China (PRC) goes out
Passport control is quarantined, and professional standard (SN/T 1870-2007) is middle to be provided: culture solution needs in centrifuge 8000r/min's
Under the conditions of be centrifuged 5min.
Automation microbial molecules detection technique in the prior art is not related to the pre-treatment step of sample, i.e. sample is pre-
Processing must be by manually having been manually done.To find out its cause, in high volume needing sample to be measured, after sample is injected porous plate,
Porous plate concussion instrument in the prior art is not suitable for cooperating with other precision instruments, because after porous plate concussion instrument earthquake usually
There are various displacement errors.It is automation microbial molecules so how to realize separation of solid and liquid of the nucleic acid cleavage liquid on porous plate
The technical bottleneck of detection technique development.
Summary of the invention
The purpose of the present invention is overcome the deficiencies of the prior art and provide a kind of nucleic acid cleavage liquid separation side based on porous plate
Method allows the sample in porous plate to be automatically performed the separation of nucleic acid cleavage liquid under the premise of not generating excessive vibration.
The solution to the technical problem of the present invention is as follows: a kind of nucleic acid cleavage liquid separating method based on porous plate, packet
Include following steps:
(1) it takes filtering porous plate and accepts porous plate, filter membrane is provided in the reacting hole of filtering porous plate and is connected to and is reacted
The channel of hole interior or exterior space；
(2) the upper building enclosure of set outside porous plate is being accepted；
(3) filtering porous plate makes building enclosure form confined space in building enclosure top cover；
(4) nucleic acid cleavage liquid is injected in the reacting hole of filtering porous plate, which can also move on to the advance of step (3)
Row；
(5) negative pressure is formed in the confined space that building enclosure is formed, makes the nucleic acid cleavage in filtering porous plate reacting hole
Liquid enters underlying undertaking porous plate through filter membrane.
As an improvement, forming the mode of negative pressure in the confined space that building enclosure is formed in the step (5) are as follows: will
Building enclosure is connected to negative pressure device and opens negative pressure device.
As an improvement, forming the mode of negative pressure in the confined space that building enclosure is formed in the step (5) are as follows: will
It accepts porous plate to place on the base, pedestal is connected to negative pressure device；Set building enclosure and filtering are added on porous plate when accepting
After porous plate, filtering porous plate, pedestal form confined space between building enclosure.
As a further improvement, the movement of the undertaking porous plate, building enclosure, filtering porous plate is by there is computer control
The mechanical arm of system is completed.
It is a further object of the present invention to provide a kind of device for realizing the above method, technical solutions are as follows:
A kind of nucleic acid cleavage liquid separating apparatus based on porous plate, including filtering porous plate, accept porous plate, go along with sb. to guard him knot
Structure, vacuum pump, be provided in the reacting hole of filtering porous plate filter membrane be connected to the channel of reacting hole interior or exterior space, accept porous plate
Inside building enclosure, filtering porous plate is located at the top of building enclosure, is formed between filtering porous plate, building enclosure closed
Space, building enclosure are connected to vacuum pump.
As an improvement, the device further includes porous plate and the filtering porous plate accepted by computer-controlled mechanical arm
Movement is completed by mechanical arm.
As alternative technical solution, which includes pedestal, filtering porous plate, accepts porous plate, is building enclosure, true
Sky pumps, and filter membrane is provided in the reacting hole of filtering porous plate and is located at the channel of reacting hole interior or exterior space, undertaking porous plate is connected to
Simultaneously be located on pedestal inside building enclosure, filtering porous plate is located at the top of building enclosure, filtering porous plate, building enclosure,
Confined space is formed between pedestal, pedestal is connected to vacuum pump.
As an improvement, the device further includes by computer-controlled mechanical arm, the undertaking porous plate, building enclosure, filtering
The movement of porous plate is completed by mechanical arm.
So-called filtering porous plate is with the difference between common porous plate in the application: filtering porous plate reacts
Channel is offered in hole, filter membrane is covered at channel, and the sample in reacting hole can flow under the action of negative pressure through filter membrane
Except reacting hole.
So-called undertaking porous plate, that is, common porous in the prior art plate in the application accepts reacting hole on porous plate
Quantity, volume, position are corresponding to the quantity, volume of reacting hole, position on filtering porous plate, are used to catch from filtering porous plate
The liquid filtered out.
The application makes the nucleic acid cleavage liquid being located in porous plate reacting hole not generate excessive vibration using porous plate as carrier
Under the premise of realize and be automatically separated, so that sample pretreatment step is incorporated into automation microbial molecules detection in the prior art and set
It is standby to be possibly realized.The application can once complete the separation of multiple samples, and lock out operation improves work effect without human error
Rate.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the embodiment of the present invention 1；
Fig. 2 is that porous plate, filtering porous plate, the assembling schematic diagram between building enclosure are accepted in the embodiment of the present invention 1；
Fig. 3 is the structural schematic diagram of the embodiment of the present invention 2；
Fig. 4 is pedestal in the embodiment of the present invention 2, accepts porous plate, filtering porous plate, the assembly signal between building enclosure
Figure；
Fig. 5 is the structural schematic diagram for the reacting hole that the present invention accepts porous plate.
In figure: 1, filtering porous plate；2, porous plate is accepted；3, building enclosure；4, vacuum pump；5, mechanical arm；6, pedestal；7,
Channel.
Specific embodiment
Embodiment 1
In the application, the nucleic acid cleavage liquid separating method based on porous plate includes the following steps:
(1) it takes filtering porous plate and accepts porous plate, filter membrane is provided in the reacting hole of filtering porous plate and is connected to and is reacted
The channel of hole interior or exterior space；
(2) the upper building enclosure of set outside porous plate is being accepted；
(3) in building enclosure top cover filtering porous plate make building enclosure formed confined space, filtering porous plate it is anti-
The reacting hole that Ying Kongyu accepts porous plate corresponds up and down；
(4) nucleic acid cleavage liquid will be injected in the reacting hole of filtering porous plate, which can also move on to the advance of step (3)
Row；
(5) negative pressure is formed in the confined space that building enclosure is formed makes the nucleic acid cleavage liquid in filtering porous plate reacting hole
Enter underlying undertaking porous plate through filter membrane.
In step (5), building enclosure formed confined space in formed negative pressure mode are as follows: by building enclosure with
Negative pressure device is connected to and opens negative pressure device.
The movement of porous plate, building enclosure, filtering porous plate is accepted by there is computer-controlled mechanical arm to complete.
Device corresponding with this method is as shown in Figs. 1-2, a kind of nucleic acid cleavage liquid separating apparatus based on porous plate, including
Filtering porous plate 1, accept porous plate 2, building enclosure 3, vacuum pump 4, by computer-controlled mechanical arm 5.Filtering porous plate 1 it is anti-
Answer hole as shown in figure 5, be provided in reacting hole filter membrane be connected to the channel 7 of reacting hole interior or exterior space.Building enclosure 3 is spacious for top
The container opened, building enclosure 3 have 4 sides and 1 bottom surface, and open top can cooperate with filtering porous plate 1.It accepts more
Orifice plate 2 is located at the inside of building enclosure 3, and filtering porous plate 1 is located at the top of building enclosure 3, filtering porous plate 1, building enclosure 3
Between form confined space, building enclosure 3 is connected to vacuum pump 4.The movement of porous plate 2, filtering porous plate 1 is accepted by mechanical arm
5 controls are completed.
Embodiment 2
Method described in the present embodiment the difference from embodiment 1 is that: in step (5), building enclosure formed it is close
It closes in space and forms the mode of negative pressure are as follows: porous plate will be accepted and placed on the base, pedestal is connected to negative pressure device；It is more when accepting
After adding set building enclosure and filtering porous plate on orifice plate, filtering porous plate, pedestal form confined space between building enclosure.
As shown in Figure 3-4 is device corresponding with this method.The device include pedestal 6, filtering porous plate 1, accept it is more
Orifice plate 2, building enclosure 3, vacuum pump 4, by computer-controlled mechanical arm 5.Be provided in the reacting hole of filtering porous plate 1 filter membrane and
It is connected to the channel 7 of reacting hole interior or exterior space, accepts the inside that porous plate 2 is located at building enclosure 3, porous plate 2 is accepted and is placed on base
On seat 6, pedestal 6 is connected to vacuum pump 4.Building enclosure 3 goes to upper and lower open structure, and top can match with filtering porous plate 1
It closes, bottom can cooperate with pedestal 6.Filtering porous plate 1 is located at the top of building enclosure 3, filtering porous plate 1, building enclosure 3,
Confined space is formed between pedestal 6, the movement for accepting porous plate 2, building enclosure 3, filtering porous plate 1 has been controlled by mechanical arm 5
At.
Claims (5)
1. a kind of nucleic acid cleavage liquid separating method based on porous plate, which comprises the steps of:
(1) take filtering porous plate and accept porous plate, be provided in the reacting hole of filtering porous plate filter membrane be connected in reacting hole
The channel of the external space；
(2) porous plate will be accepted to be placed on the pedestal being connected to negative pressure device；
(3) the upper building enclosure of set outside porous plate is being accepted, building enclosure goes to upper and lower open structure；
(4) filtering porous plate makes filtering porous plate, pedestal, forms confined air between building enclosure in building enclosure top cover
Between；
(5) nucleic acid cleavage liquid is injected in the reacting hole of filtering porous plate；
(6) negative pressure is formed in the confined space that building enclosure is formed, keeps the nucleic acid cleavage liquid in filtering porous plate reacting hole saturating
Filter membrane enters underlying undertaking porous plate.
2. a kind of nucleic acid cleavage liquid separating method based on porous plate, which comprises the steps of:
(1) take filtering porous plate and accept porous plate, be provided in the reacting hole of filtering porous plate filter membrane be connected in reacting hole
The channel of the external space；
(2) porous plate will be accepted to be placed on the pedestal being connected to negative pressure device；
(3) the upper building enclosure of set outside porous plate is being accepted, building enclosure goes to upper and lower open structure；
(4) nucleic acid cleavage liquid is injected in the reacting hole of filtering porous plate；
(5) filtering porous plate makes filtering porous plate, pedestal, forms confined air between building enclosure in building enclosure top cover
Between；
(6) negative pressure is formed in the confined space that building enclosure is formed, keeps the nucleic acid cleavage liquid in filtering porous plate reacting hole saturating
Filter membrane enters underlying undertaking porous plate.
3. the nucleic acid cleavage liquid separating method based on porous plate as claimed in claim 1 or 2, which is characterized in that described holds
The movement of porous plate, building enclosure, filtering porous plate is connect by there is computer-controlled mechanical arm to complete.
4. a kind of nucleic acid cleavage liquid separating apparatus based on porous plate, which is characterized in that including pedestal, filtering porous plate, accept
Porous plate, building enclosure, vacuum pump, be provided in the reacting hole of filtering porous plate filter membrane be connected to the logical of reacting hole interior or exterior space
Road, building enclosure go to upper and lower open structure, accept porous plate and are located inside building enclosure and are located on pedestal, filter more
Orifice plate is located at the top of building enclosure, and filtering porous plate, building enclosure form confined space, pedestal and vacuum pump between pedestal
Connection；Accept quantity, volume, quantity, volume, the position phase of position and reacting hole on filtering porous plate of reacting hole on porous plate
It answers, accepts porous plate for catching the liquid filtered out from filtering porous plate.
5. the nucleic acid cleavage liquid separating apparatus based on porous plate as claimed in claim 4, which is characterized in that further include by computer
The mechanical arm of control, the movement for accepting porous plate, building enclosure, filtering porous plate are completed by mechanical arm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610579824.8A CN106190817B (en) | 2016-07-19 | 2016-07-19 | Nucleic acid cleavage liquid separating method and device based on porous plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610579824.8A CN106190817B (en) | 2016-07-19 | 2016-07-19 | Nucleic acid cleavage liquid separating method and device based on porous plate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106190817A CN106190817A (en) | 2016-12-07 |
| CN106190817B true CN106190817B (en) | 2019-01-08 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610579824.8A Active CN106190817B (en) | 2016-07-19 | 2016-07-19 | Nucleic acid cleavage liquid separating method and device based on porous plate |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003091407A2 (en) * | 2002-04-24 | 2003-11-06 | Hitachi Chemical Research Center, Inc. | Device and method for high-throughput quantification of mrna from whole blood |
| EP1997912A2 (en) * | 2003-10-30 | 2008-12-03 | Hitachi Chemical Company, Ltd. | Device and method for high-throughput quantification of MRNA from whole blood |
| CN205803485U (en) * | 2016-07-19 | 2016-12-14 | 湖州高亿诺生物科技有限公司 | Nucleic acid cleavage liquid separating apparatus based on porous plate |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080113357A1 (en) * | 2006-06-29 | 2008-05-15 | Millipore Corporation | Filter device for the isolation of a nucleic acid |
-
2016
- 2016-07-19 CN CN201610579824.8A patent/CN106190817B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003091407A2 (en) * | 2002-04-24 | 2003-11-06 | Hitachi Chemical Research Center, Inc. | Device and method for high-throughput quantification of mrna from whole blood |
| EP1997912A2 (en) * | 2003-10-30 | 2008-12-03 | Hitachi Chemical Company, Ltd. | Device and method for high-throughput quantification of MRNA from whole blood |
| CN205803485U (en) * | 2016-07-19 | 2016-12-14 | 湖州高亿诺生物科技有限公司 | Nucleic acid cleavage liquid separating apparatus based on porous plate |
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|---|---|
| CN106190817A (en) | 2016-12-07 |
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Effective date of registration: 20190419 Address after: 313000 Eleven Floors of H Building, Freeport, Headquarters, Eastern New Town, Huzhou City, 1188 Fulu Road, Wuxing District, Huzhou City, Zhejiang Province Patentee after: Huzhou billion Connaught Biological Technology Co., Ltd. Address before: 325000 Room 201, 2nd Floor, Technology Business Incubator Pilot Building, Venture Service Center, Wenzhou High-tech Industrial Development Zone, Zhejiang Province (Independent Declaration) Patentee before: Wenzhou Gaoyino Biotechnology Co., Ltd. |
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Effective date of registration: 20200824 Address after: 1213-177, 12 / F, building 3, No. 1366, Hongfeng Road, Kangshan street, Huzhou Economic and Technological Development Zone, Huzhou City, Zhejiang Province Patentee after: Original microbial detection technology (Huzhou) Co., Ltd Address before: 313000 Eleven Floors of H Building, Freeport, Headquarters, Eastern New Town, Huzhou City, 1188 Fulu Road, Wuxing District, Huzhou City, Zhejiang Province Patentee before: HUZHOU GAOYINUO BIOLOGICAL TECHNOLOGY Co.,Ltd. |
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