CN218879913U

CN218879913U - Automatic device for extracting plasmid DNA by alkaline lysis

Info

Publication number: CN218879913U
Application number: CN202222573512.XU
Authority: CN
Inventors: 崔林林; 谢德秀; 孙锋; 朱晨曦
Original assignee: Suzhou Ansi Microbial Technology Co ltd
Current assignee: Suzhou Ansi Microbial Technology Co ltd
Priority date: 2022-09-28
Filing date: 2022-09-28
Publication date: 2023-04-18
Anticipated expiration: 2032-09-28

Abstract

The utility model relates to an automation equipment of plasmid DNA is drawed in alkali schizolysis, including the stoste pipeline, alkali lye pipeline, in the dynamic and static mixing tank neutralization liquid pipeline, the vibration mixes the pond, magnetic stirring container, deep filtration membrane and collection container, dynamic and static mixing tank upper end is equipped with a dynamic mixing district, the one end department in dynamic mixing district is equipped with stoste pipeline and alkali lye pipeline, and other end department is equipped with a static mixing district, the vibration mixes the pond and is equipped with a driving motor and oscillating cam, neutralization liquid pipeline, the liquid outlet mixes pond fixed connection with the vibration respectively, concatenate the PH sensor between vibration mixing tank and the magnetic stirring container, first valve, concatenate the motor pump between magnetic stirring container and the collection container, the deep filtration membrane. This automation equipment of plasmid DNA is drawed in alkali lysis has and makes things convenient for the dismouting and changes the part, improves and mixes the degree of consistency, improves advantages such as filter effect and yield.

Description

Automatic device for extracting plasmid DNA by alkaline lysis

Technical Field

The utility model relates to an alkali schizolysis draws plasmid DNA technical field in the bio-pharmaceuticals, especially an automatic equipment that alkali schizolysis draws plasmid DNA.

Background

The alkaline cracking method is a method for preparing plasmid DNA with the most extensive application, and achieves the purpose of denaturing and extracting the plasmid DNA by adding alkaline cracking liquid under an escherichia coli or yeast culture system. The principle is based on the difference between the denaturation and renaturation of chromosomal DNA and plasmid DNA. Under alkaline conditions of pH, the hydrogen bonds of chromosomal DNA are broken and the double helix structure is released and denatured. Most hydrogen bonds of plasmid DNA are broken, but two complementary strands of double-helix covalent closed circular are not completely separated, when the pH value of the plasmid DNA is adjusted to be neutral by a high-salt buffer solution with pH4.8, the denatured plasmid DNA restores the original configuration and is stored in the solution, but the chromosome DNA cannot be renatured to form a winding net structure, a turbid harvest solution is obtained at the moment, and the harvest solution is filtered, so that the stable and high-yield plasmid DNA can be obtained. The invention mainly solves the problems that the mixing of lysate and a sample is not sufficient in the mixing process, the contact of a medium is not complete, the process is not controllable, the shearing force in the large process destroys plasmid DNA, and the disposable use requirement cannot be met.

The simple mixing mode is that firstly, the sample and the lysis solution are not mixed fully, and the static mixing tube cannot make the medium contact completely. Secondly, the process is uncontrollable, the mixing degree cannot be actively controlled only by static mixing or atomization, and no means for implementing intervention control is available. Then, the mixture is statically mixed at a high flow rate, the liquid has a great shearing force, and the atomized mixture can directly damage the topological structure of the plasmid DNA, so that the plasmid DNA is denatured or difficult to renature. And finally, static mixing and atomization mixing cannot meet the requirement of one-time use, because the static mixing pipe and the flow path are integrated and are difficult to replace, and the atomizer is formed by equipment hardware and cannot be replaced.

Therefore, we developed an automatic device for extracting plasmid DNA by alkaline lysis, which is used to solve the above problems.

SUMMERY OF THE UTILITY MODEL

The utility model aims at overcoming prior art's not enough and provide an alkaline lysis and draw automation equipment of plasmid DNA, have and make things convenient for the dismouting and change the part, improve the mixed degree of consistency, improve advantages such as filter effect and collection rate.

In order to achieve the above purpose, the utility model adopts the technical scheme that: the utility model provides an automatic equipment of plasmid DNA is drawed in alkali schizolysis, includes that an adjustable stoste pipeline, an adjustable alkali lye pipeline, a dynamic and static mix pond, an adjustable neutralization liquid pipeline, an oscillation mix pond, a magnetic stirring container, a deep filtration membrane and a collection container, the one end in dynamic and static mix pond respectively with stoste pipeline alkali lye pipeline fixed connection to other end department is equipped with a liquid outlet, neutralization liquid pipeline the liquid outlet respectively with oscillation mix pond fixed connection, oscillation mix the pond with concatenate a PH sensor, a first valve between the magnetic stirring container, the magnetic stirring container with collect concatenate an electric pump between the container the deep filtration membrane.

Preferably, the oscillation mixing pool is provided with a first driving motor, the first driving motor is fixedly connected with an oscillation cam, and the oscillation cam is abutted to a support.

Preferably, the support is of a T-shaped structure, the support is provided with a stand column, and the stand column is wound with a lengthened hose.

Preferably, the dynamic and static mixing tank upper end is equipped with a dynamic mixing area, the one end department in dynamic mixing area is equipped with stoste pipeline with alkali lye pipeline to the other end department is equipped with a static mixing area, is close to alkali lye pipeline one side department is equipped with an inside rotor, static mixing area with the liquid outlet intercommunication.

Preferably, a cavity is arranged at the lower end of the dynamic and static mixing pool, a second driving motor is arranged in the cavity and connected to the external magnetic rotor through a connecting piece, and the external magnetic rotor is arranged on the lower side of the internal rotor.

Preferably, the connecting member is a synchronous pulley or a gear set.

Preferably, the stock solution conveying pipeline is sequentially provided with a first conveying pump, a first pressure sensor and a first flowmeter, and the first flowmeter is arranged close to the dynamic and static mixing pool.

Preferably, the alkali liquor conveying pipeline is sequentially provided with a second conveying pump, a second pressure sensor and a second flowmeter, and the second flowmeter is arranged close to the dynamic and static mixing pool.

Preferably, the PH sensor is connected to the waste liquid tank through a second valve.

Preferably, the neutralizing liquid conveying pipeline is sequentially provided with a third conveying pump, a third pressure sensor and a third flowmeter, and the liquid outlet and the neutralizing liquid conveying pipeline are respectively connected with the lengthened hose.

Because of above-mentioned technical scheme's application, compared with the prior art, the utility model have the following advantage:

the automatic device for extracting plasmid DNA by alkaline cracking of the utility model comprises a dynamic and static mixing tank, which can fully and uniformly stir stock solution and alkali liquor, and the dynamic and static mixing can be completed in the same mixing tank, thus improving the cracking mixing uniformity; the oscillating mixing pool is adjusted by a cam to improve the neutralization mixing uniformity; after magnetic stirring, the mixture is filtered by a deep filtration membrane, so that the filtration effect and the yield are improved; automatic system's equipment makes things convenient for equipment dismouting and change.

Drawings

FIG. 1 is a schematic structural diagram of an automatic apparatus for extracting plasmid DNA by alkaline lysis.

Figure 2 is the utility model discloses a structure schematic diagram of sound attitude mixing tank.

Figure 3 is the structure schematic diagram of the oscillating mixing tank of the utility model.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

In the attached fig. 1 to 3, an automatic apparatus for extracting plasmid DNA by alkaline lysis comprises an adjustable stock solution delivery pipe 19, an adjustable alkali solution delivery pipe 20, a dynamic and static mixing tank 10, an adjustable neutralizing solution delivery pipe 301, an oscillating mixing tank 11, a magnetic stirring container 15, a deep filtration membrane 17 and a collection container 18. The neutralizing liquid conveying pipeline 301 is sequentially provided with a third conveying pump 3, a third pressure sensor 6 and a third flow meter 9, the liquid outlet 27 and the neutralizing liquid conveying pipeline 301 are respectively connected with the lengthened hose 28, the stock solution and the alkali liquor are subjected to alkali cracking reaction in the dynamic and static mixing tank 10, and the reacted solution and the neutralizing liquid are uniformly mixed in the oscillating and mixing tank 11.

In order to control the mixing ratio of the stock solution and the alkali solution, one end of the dynamic and static mixing tank 10 is fixedly connected with a stock solution conveying pipeline 19 and an alkali solution conveying pipeline 20 respectively. In order to monitor the pressure flow of the stock solution, the stock solution conveying pipeline 19 is sequentially provided with a first conveying pump 1, a first pressure sensor 4 and a first flow meter 7, and the first flow meter 7 is arranged close to the dynamic and static mixing tank 10. In order to monitor the pressure flow of the alkali liquor, the alkali liquor conveying pipeline 20 is sequentially provided with a second conveying pump 2, a second pressure sensor 5 and a second flowmeter 8, and the second flowmeter 8 is arranged close to the dynamic and static mixing tank 10.

A liquid outlet 27 is arranged at the other end of the dynamic and static mixing tank 10, the neutralizing liquid conveying pipeline 301 and the liquid outlet 27 are respectively and fixedly connected with the oscillating mixing tank 11, a PH sensor 12 and a first valve 13 are connected in series between the oscillating mixing tank 11 and the magnetic stirring container 15, and the PH sensor 12 and the first valve 13 are connected to the waste liquid tank 14 through a second valve. An electric pump 16 and a deep filtration membrane 17 are connected in series between the magnetic stirring container 15 and the collection container 18, the mixed liquid meeting the collection requirement is filtered by the deep filtration membrane 17, and the unsatisfied mixed liquid is discharged to the waste liquid tank 14.

In order to improve the mixing uniformity, the oscillating mixing tank 11 is integrally composed of an elongated hose 28, a hose bracket 29, an oscillating cam 30 and a first driving motor 31, wherein the elongated hose 28 is optimally sized to 15 meters and 30 meters according to relevant test data.

The oscillating mixing tank 11 is provided with a first driving motor 31, the first driving motor 31 is fixedly connected with an oscillating cam 30, and the oscillating cam 30 is abutted to a bracket 29. The bracket 29 is of a T-shaped structure, the bracket 29 is provided with a stand column 291, and the stand column 291 is wound with an elongated hose 28. The first driving motor 31 drives the oscillating cam 30 to drive the upright 291 to oscillate, so that the mixed liquid in the elongated hose 28 is fully and uniformly mixed.

Wherein the interior of the dynamic and static mixing pool 10 is composed of a stock solution conveying pipeline 19, an alkali liquor conveying pipeline 20, a dynamic mixing area 21, an inner rotor 22, a static mixing area 23, an outer magnetic rotor 24, a synchronous belt wheel or a gear set, a second driving motor 26 and a liquid outlet 27. The function of the device is to efficiently assist the mixing of two liquids.

The upper end of the dynamic and static mixing pool 10 is provided with a dynamic mixing area 21, one end of the dynamic mixing area 21 is provided with a stock solution conveying pipeline 19 and an alkali liquor conveying pipeline 20, the other end is provided with a static mixing area 23, one side close to the alkali liquor conveying pipeline 20 is provided with an inner rotor 22, and the static mixing area 23 is communicated with a liquid outlet 27.

In order to uniformly mix the alkali cracking, a chamber 101 is arranged at the lower end of the dynamic and static mixing tank 10, a second driving motor 26 is arranged in the chamber 101, the second driving motor 26 is connected to the external magnetic rotor 24 through a connecting piece 25, and the connecting piece 25 is a synchronous pulley or a gear set in order to improve the convenience of installation. The external magnetic rotor 24 is arranged at the lower side of the internal rotor 22, and the external magnetic rotor 24 drives the internal rotor 22 to stir the mixed liquid of the stock solution and the alkali solution.

Stock solution and alkali liquor are synchronously pumped into a dynamic and static mixing pool 10 through a first delivery pump 1 and a second delivery pump 2 respectively, pressure states are monitored through a first pressure sensor 4 and a second pressure sensor 5 respectively, and flow rate conditions of the pumps are monitored through a first flow meter 7 and a second flow meter 8 respectively. The mixed reaction liquid enters the oscillation mixing pool 11 through a pipeline, synchronous neutralization liquid is pumped into the oscillation mixing pool 11 by the third delivery pump 3, the neutralization liquid is monitored by the third pressure sensor 6 at the moment, and the flow rate is monitored by the third flow meter 9.

A PH sensor 12 is arranged at the rear end of the oscillation mixing pool, and whether the mixed liquid meets the collection requirement or not is judged according to the PH value of the final mixed liquid. If the condition is satisfied, the waste liquid is collected by the magnetic stirring container 15 at the end of the first valve 13, and if the condition is not satisfied, the waste liquid is directly discharged to the waste liquid tank 14 from the end of the second valve for treatment. And the solution meeting the PH value is obtained by continuously mixing the solution with the magnetic stirring container 15 and then pumping the mixture into a deep filtration membrane 17 by an electric pump 16 for filtration.

The above is only a specific application example of the present invention, and does not set any limit to the protection scope of the present invention. All the technical solutions formed by equivalent transformation or equivalent replacement fall within the protection scope of the present invention.

Claims

1. An automatic equipment for extracting plasmid DNA by alkaline lysis is characterized in that: including an adjustable stoste pipeline (19), an adjustable alkali liquor pipeline (20), a static mixing tank (10), an adjustable neutralization liquid pipeline (301), an oscillation mixing tank (11), a magnetic stirring container (15), a deep filtration membrane (17) and a collection container (18), the one end of static mixing tank (10) respectively with stoste pipeline (19), alkali liquor pipeline (20) fixed connection to the other end department is equipped with a liquid outlet (27), neutralization liquid pipeline (301), liquid outlet (27) respectively with oscillation mixing tank (11) fixed connection, oscillation mixing tank (11) with concatenate a PH sensor (12), a first valve (13) between magnetic stirring container (15), magnetic stirring container (15) with collect and concatenate an electric pump (16) between container (18), deep filtration membrane (17).

2. The automated alkaline lysis plasmid DNA extraction apparatus of claim 1, wherein the oscillation mixing tank (11) is provided with a first driving motor (31), the first driving motor (31) is fixedly connected with an oscillation cam (30), and the oscillation cam (30) is abutted to a bracket (29).

3. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 2, wherein the support (29) is a T-shaped structure, the support (29) is provided with a column (291), and the column (291) is wound with an elongated hose (28).

4. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 1, wherein the dynamic and static mixing tank (10) is provided with a dynamic mixing zone (21) at the upper end, the raw liquid conveying pipe (19) and the alkaline liquid conveying pipe (20) are provided at one end of the dynamic mixing zone (21), and a static mixing zone (23) is provided at the other end, an inner rotor (22) is provided at one side close to the alkaline liquid conveying pipe (20), and the static mixing zone (23) is communicated with the liquid outlet (27).

5. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 4, wherein the dynamic and static mixing tank (10) is provided with a chamber (101) at the lower end, the chamber (101) is provided with a second driving motor (26), the second driving motor (26) is connected to the outer magnetic rotor (24) through a connecting member (25), and the outer magnetic rotor (24) is arranged at the lower side of the inner rotor (22).

6. The automated apparatus for alkaline lysis extraction of plasmid DNA according to claim 5, characterized in that the connection piece (25) is a synchronous pulley or a gear train.

7. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 1, wherein the stock solution delivery pipe (19) is sequentially provided with a first delivery pump (1), a first pressure sensor (4) and a first flow meter (7), and the first flow meter (7) is disposed near the dynamic-static mixing tank (10).

8. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 1, wherein the alkaline solution delivery pipe (20) is sequentially provided with a second delivery pump (2), a second pressure sensor (5) and a second flow meter (8), and the second flow meter (8) is disposed near the dynamic and static mixing tank (10).

9. The automated apparatus for alkaline lysis extraction of plasmid DNA according to claim 1, characterized in that the space between the pH sensor (12) and the first valve (13) is connected to a waste liquid tank (14) via a second valve.

10. The automated alkaline lysis plasmid DNA extraction apparatus according to claim 3, wherein the neutralization solution delivery conduit (301) is sequentially provided with a third delivery pump (3), a third pressure sensor (6) and a third flow meter (9), and the liquid outlet (27) and the neutralization solution delivery conduit (301) are respectively connected with the elongated hose (28).