Alkali cracking device

Abstract

The utility model relates to the technical field of escherichia coli cracking equipment, and particularly discloses an alkali cracking device which comprises a container, a stirring paddle and a motor, the stirring paddle is rotatably arranged in the container; the motor is in transmission connection with the stirring paddle and is used for driving the stirring paddle to rotate. According to the alkali cracking device provided by the invention, the solution can be uniformly mixed and more plasmids can be released in effective time in the production of an escherichia coli alkali cracking process, so that a harvest solution with higher clarity and better plasmid quality is obtained, and the effects of improving the production efficiency and the quality stability are achieved.

Description

Technical Field

[0001] The present application relates to the technical field of Escherichia coli lysis equipment, and in particular to an alkaline lysis device. Background Art

[0002] Alkaline lysis is one of the commonly used methods for E. coli lysis due to its wide applicability and good repeatability. In small-scale plasmid extraction, the existing method of alkaline lysis of E. coli is usually to achieve gentle mixing and lysis by hand-cranking or stirring with a handheld blender. However, the mixing speed, angle and strength control of different operators are inconsistent, and the same operator is also difficult to control the stability of different batches, and the production efficiency is low. Utility Model Content

[0003] In view of this, the purpose of the present application is to provide an alkaline lysis device to solve the problem that the existing alkaline lysis method for Escherichia coli has low production efficiency and unstable quality of different batches.

[0004] In order to achieve the above technical purpose, the present application provides an alkaline cracking device, including: a container, a stirring paddle and a motor;

[0005] The stirring paddle is rotatably disposed in the container;

[0006] The motor is drivingly connected to the stirring paddle to drive the stirring paddle to rotate.

[0007] Furthermore, the ratio of the diameter of the stirring paddle to the inner diameter of the container is 0.25 to 0.5.

[0008] Furthermore, the stirring paddle includes an axis located in the middle, a radial portion extending in the radial direction, and two axial portions extending in the axial direction;

[0009] The axis, the radial portion and the two axial portions constitute an anchor-type paddle.

[0010] Furthermore, the ratio of the axial height of the axial portion to the diameter of the stirring paddle is 0.8 to 1.2.

[0011] Further, the radial width of the axial portion is 0.1 to 0.2 times the diameter of the stirring paddle;

[0012] The axial height of the radial portion is 0.1 to 0.2 times the diameter of the stirring paddle;

[0013] The diameter of the axis is 0.08 to 0.2 times the diameter of the stirring paddle.

[0014] Further, an axial extension portion is connected between the radial portion and the axis;

[0015] The axial extension portion is used to increase the connection area between the radial portion and the axis.

[0016] Further, the axial height of the axial extension portion is ≤ 1 / 3 of the height of the axial portion;

[0017] The radial width of the axially extending portion is ≤ 1 / 4 of the diameter of the stirring paddle.

[0018] Furthermore, the stirring paddle includes a plurality of paddle blades evenly distributed around the circumference, and the paddle blades are arranged at an angle.

[0019] Furthermore, the stirring propeller is a twin-screw propeller.

[0020] Furthermore, the stirring propeller is a single propeller.

[0021] Furthermore, the ratio of the height of the stirring paddle along the axial direction to the diameter of the stirring paddle is 0.8 to 1.2.

[0022] Furthermore, the ratio of the height to the inner diameter of the container is 1 to 1.5.

[0023] Further, the ratio of the height to the inner diameter of the container is greater than 1.5;

[0024] The stirring paddle is provided with a plurality of paddle blades at intervals along the axial direction.

[0025] Further, it also includes: bagging;

[0026] The bag is sleeved in the container;

[0027] The stirring paddle extends into the bag.

[0028] Further, it also includes: a diaphragm;

[0029] The bag is an open bag;

[0030] The membrane is placed in the bag to cover the top of the bag;

[0031] The diaphragm is hung in the container;

[0032] The stirring paddle passes through the membrane and extends into the bag.

[0033] Furthermore, the bag is a retractable bag with a tightening rope at the bag opening.

[0034] It can be seen from the above technical scheme that the present application provides an alkaline lysis device, including: a container, a stirring paddle and a motor; the stirring paddle is rotatably arranged in the container; the motor is transmission-connected to the stirring paddle to drive the stirring paddle to rotate.

[0035] The alkaline lysis device provided in the present application can mix the solution within an effective time in the alkaline lysis process of Escherichia coli to release more plasmids and obtain a harvest liquid with higher clarity and better plasmid quality, thereby improving production efficiency and quality stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0037] Figure 1 A schematic diagram of the overall structure of an alkaline lysis device provided in an embodiment of the present application;

[0038] Figure 2 A wireframe schematic diagram of an alkaline lysis device provided in an embodiment of the present application;

[0039] Figure 3 A wireframe schematic diagram of a stirring paddle in an alkaline cracking device provided in an embodiment of the present application having multiple blades arranged along the axial direction;

[0040] Figure 4 A schematic diagram of an alkaline cracking device provided in an embodiment of the present application in which a stirring paddle is a multi-blade paddle;

[0041] Figure 5 A schematic diagram of a stirring paddle in a mountain-shaped agitator in an alkaline cracking device provided in an embodiment of the present application;

[0042] Figure 6 A schematic diagram of an alkaline cracking device provided in an embodiment of the present application in which the stirring propeller is a double propeller;

[0043] Figure 7 A schematic diagram of a stirring propeller as a single propeller in an alkaline cracking device provided in an embodiment of the present application;

[0044] Figure 8 A schematic diagram of a bagging device in an alkaline lysis device provided in an embodiment of the present application;

[0045] Fig. 9 A schematic diagram of a membrane in an alkaline lysis device provided in an embodiment of the present application;

[0046] Fig.10 A schematic diagram of an open-mouth bag in an alkaline lysis device provided in an embodiment of the present application. DETAILED DESCRIPTION

[0047] The technical solutions of the embodiments of the present application will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the specification of the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection requested by the present application.

[0048] In the description of the embodiments of the present application, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the embodiments of the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the embodiments of the present application. In addition, the terms "first", "second", and "third" are used for descriptive purposes only and cannot be understood as indicating or implying relative importance.

[0049] In the description of the embodiments of the present application, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a replaceable connection, or an integral connection, it can be a mechanical connection, it can be an electrical connection, it can be a direct connection, it can be indirectly connected through an intermediate medium, and it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the embodiments of the present application can be understood according to specific circumstances.

[0050] See also Figure 1 An alkaline lysis device provided in an embodiment of the present application includes: a container 10, a stirring paddle 20 and a motor 30; the stirring paddle 20 is rotatably arranged in the container 10; the motor 30 is transmission-connected to the stirring paddle 20, and is used to drive the stirring paddle 20 to rotate.

[0051] By using the alkaline lysis device provided in this embodiment, when performing alkaline lysis of Escherichia coli, constant-speed and timed stirring of the solution can be achieved, thereby improving production efficiency and ensuring the stability of production quality of different batches.

[0052] In a further improved embodiment, the ratio of the diameter d of the stirring paddle 20 to the inner diameter D of the container 10 is 0.25 to 0.5.

[0053] Specifically, the inventors have found that in practical applications, the ratio between the inner diameter of the container 10 and the diameter of the stirring paddle 20 has a great influence on the mixing efficiency. In this solution, setting d / D to 0.25 to 0.5 can achieve better mixing efficiency.

[0054] In one embodiment, see Figures 1 to 4In this embodiment, the stirring paddle 20 is an anchor-type paddle.

[0055] In practical applications, the stirring paddle 20 can stir the solution in the radial direction by rotating, but stirs less in the vertical direction, that is, in the axial direction; on the one hand, it is necessary to consider the distance between the liquid surface and the top of the container required when a large number of bubbles are generated by adding flocculants after neutralization, and on the other hand, it is necessary to consider the height at which the paddle 21 can complete the mixing when the paddle 21 in the stirring paddle 20 is fixed. For this reason, the anchor paddle used as the stirring paddle 20 in this embodiment can improve the stirring of the stirring paddle 20 in the axial direction, thereby improving the stirring effect.

[0056] As an anchor-type impeller, in this embodiment, the ratio of the axial height h to the diameter d of the impeller 20 is 0.8 to 1.2. Figure 2 The stirring blade 20 has two blades 21 in an L-shape. The two blades 21 are symmetrically arranged to form a mountain shape.

[0057] Specifically, the stirring paddle 20 may include an axis 214, an axial portion 211 and a radial portion 212, wherein the axis 214 is located in the middle; the radial portion 212 is arranged at the bottom of the axis 214 and extends to both sides along the radial direction of the axis 214; the axial portion 211 includes two symmetrically arranged on both sides of the radial portion 212 and perpendicular to the radial portion 212; the axis 213, the radial portion 212 and the two axial portions 211 form a mountain-shaped stirring paddle 20. Among them, the paddle 21 is composed of the axial portion 211 and the radial portion 212.

[0058] See also Figure 5 In this embodiment, the radial width a of the axial portion 211 is specifically 0.1 to 0.2 times the diameter d of the stirring paddle 20; wherein the diameter d of the stirring paddle 20 is also the radial length of the radial portion 212. The axial height b of the radial portion 212 is 0.1 to 0.2 times the diameter d. In addition, the diameter of the axis 213 is 0.08 to 0.2 times the diameter d, and the thickness of the blade 21 is 0.01 to 0.03 times the diameter d. Through the above-mentioned size restrictions, the stirring paddle 20 can have a good stirring effect while having a lighter weight.

[0059] An axial extension portion 213 is formed between the radial portion 212 and the axis 214 of the stirring paddle 20; the axial extension portion 213 is used to increase the connection area between the radial portion 212 and the axis 214, and at the same time forms a shielding portion, which can improve the firmness of the connection between the radial portion 212 and the axis 214 and improve the stirring effect. Among them, the axial height c of the axial extension portion 213 is less than or equal to 1 / 3 of the height h of the axial portion 211; the radial width e of the axial extension portion 213 is less than or equal to 1 / 4 of d, and can be 0.1 to 0.2 times the diameter d.

[0060] In another embodiment, see Figure 4 The stirring paddle 20 includes a plurality of paddle blades 21 evenly distributed around the circumference, and the paddle blades 21 are arranged at an angle.

[0061] That is, in this embodiment, the stirring paddle 20 is a multi-blade paddle, for example, a three-blade paddle or a four-blade paddle; the paddle blade 21 can be circular, elliptical or rectangular, etc. In this embodiment, the overall height h of the multi-blade paddle can be 0.3 to 0.5 times of d; the width of each paddle blade 21 can be 0.1 to 0.45 times of d, and the thickness is 0.01 to 0.03 times of d. The inclination angle of each paddle blade 21 is 30 to 60°.

[0062] In another embodiment, see Figure 6 The stirring paddle 20 can be a double propeller, the overall height h' is 0.8 to 1.2d, and a plurality of blades 21 spirally wrapped around each other in two directions can be provided. The width f of a single blade 21 is 0.12 to 0.24d, the thickness is 0.01 to 0.03d, the whole is coiled 1 to 3 times, and the coiling angle is 20 to 50°. The diameter of the stirring rod of the stirring paddle 20 is 0.08 to 0.2 times d. The inner periphery of the stirring paddle 20 is connected to the stirring rod and a blade 21 can be provided, which is spirally coiled along the stirring rod, with a width of 0.1-0.2d, a thickness of 0.01-0.03d, and a coiling angle of 50-70°. The paddle provided in this embodiment can be used for mixing high-viscosity liquids.

[0063] In another embodiment, see Figure 7 The stirring paddle 20 is a single propeller, the overall height h" is 0.8 to 1.2d, the width f' of a single blade 21 is 0.12 to 0.24d, the thickness is 0.01 to 0.03d, and the diameter of the stirring rod is 0.08 to 0.2 times of d. The upper and lower parts of the blades are basically parallel, and 3-4 blades 21 can be provided and all spirally wound in the same direction; the blade 21 is coiled 1 / 4-1 / 3 of a circle; taking the propeller provided in this embodiment as an example, the first blade 21 intersects with the bottom end of the second blade 21 at the position where the bottom end is connected to the axis 214, the second blade 21 intersects with the bottom end of the third blade 21 at the position where the bottom end is connected to the axis 214, and the third blade 21 intersects with the top bottom end of the first blade 21 at the position where the bottom end is connected to the axis 214. A spiral shape is thus formed. The angle between the side surface of the spiral single leaf and the bottom is 60-80°.

[0064] In the above embodiment, the ratio of the height to the inner diameter of the container 10 may be 1 to 1.5; and the liquid level in the container 10 may be controlled at 0.2 to 0.75H, where H is the height of the container 10 .

[0065] In practical applications, see Figure 3When the ratio of the height to the inner diameter of the container 10 is greater than 1.5, the stirring paddle 20 may be provided with a plurality of paddles 21 at intervals along the axial direction to ensure that the feed liquid can be mixed evenly.

[0066] The alkaline lysis device provided in the embodiment of the present application can be applied to stirring systems of different capacities. For example, when applied to a container 10 with a capacity of 100L, a stirring paddle 20 with a double-blade structure can be used; the connection of the blades can be fixed with screws, so that the blades are difficult to disassemble, and the stirring paddle 20 can be raised and lowered to raise the blades to different height positions of the container 10. When applied to a container 10 with a capacity of 300L or 500L, a stirrer 20 with a single-blade structure can be used, and the blades are connected to the stirring paddle 20 in a detachable manner, so as to facilitate the removal and replacement of the blades; at the same time, a weighing module, a processing module and a detection module can be provided in the container 10; the detection module can be used to detect the pH value, conductivity value and temperature value of the liquid in the container 10.

[0067] In actual application, a glass beaker is used as the container 10 for simulated stirring, and experiments are carried out using the single-propeller-shaped stirring paddle 20, the double-propeller-shaped stirring paddle 20, the mountain-shaped stirring paddle 20, the multi-blade paddle-shaped stirring paddle 20 and the existing crescent-shaped stirring paddle in the above-mentioned embodiments.

[0068] The test process is as follows: E. coli produced in the same batch is resuspended and poured into a 5L glass beaker, the bottom of the paddle is adjusted to 1 / 2 of the height of the resuspended bacterial solution, and the paddle is in the center of the beaker, and the speed is set to 200 rpm for 5 minutes of stirring and mixing.

[0069] After the stirring and mixing process, keep the blade and speed unchanged, and use a peristaltic pump to continuously add the lysis solution to the resuspended bacterial solution for 5 minutes and continue to stir and mix for 6 minutes. After that, keep the blade position and speed unchanged, use a peristaltic pump to continuously add the neutralizing solution for 5 minutes and continue to stir and mix for 6 minutes. Then, keep the blade position and speed unchanged, add 5g / L flocculant NaHCO3, stir and mix for 0.5 minutes, and finally pass through a layer of 160 mesh screen to obtain a liquid without floccules at the bottom.

[0070] According to the test results, the crescent paddle stirs gently, but the mixing efficiency is insufficient, the DNA yield is relatively low, and there are relatively more impurities remaining; the single propeller stirs violently, the supercoil ratio is relatively low, but the yield is the highest; overall, except for the crescent paddle, the other four blades can complete mixing in a shorter time.

[0071] In one embodiment, see Figure 1 and Fig.10, further comprising: a cover bag 40; the cover bag 40 is sleeved in the container 10; and the stirring paddle 20 extends into the cover bag 40. The cover bag 40 can be an existing cover bag structure.

[0072] The bag 40 is arranged in the container 10, so that the stirring paddle 20 can be directly extended into the bag 40 for stirring, and after the stirring is completed, the stirring paddle 20 is removed and the bag 40 is taken out, so that the stirred material can be completely separated from the container 10 for the next stirring operation. The bag 40 can reduce the time for cleaning the container 10, thereby reducing the production time.

[0073] In a further improved embodiment, see Fig. 9 and Fig.10 , further comprising: a diaphragm 50; the bag 40 is an open bag; the diaphragm 50 is loaded into the bag 40 to cover the top of the bag 40; the diaphragm 50 is hung in the container 10; the stirring paddle 20 passes through the diaphragm 50 and extends into the bag 40.

[0074] Specifically, unlike the existing bagging, the bagging 40 in this embodiment is an open bag, which can be easily installed and can be used in conjunction with the diaphragm 50. The outer periphery of the diaphragm 50 is provided with a hanging hole 51, which can be hung with the container 10. The diaphragm 50 can make the bagging 40 detachably connected to the container 10, and can also cover the top of the bagging 40 to prevent the material from splashing out of the bagging 40 during the stirring process.

[0075] In application, the diaphragm 50 is provided with a feed inlet 52, a stirring hole 53 and a liquid adding hole 54. The feed inlet 52 is used to add materials; the stirring hole 53 is located in the middle of the diaphragm 50 for the stirring paddle 20 to pass through; the liquid adding hole 54 can be connected to a liquid adding rod for adding liquid.

[0076] In a further improved embodiment, see Figure 8 The bag 40 is a retractable bag with a tightening rope 41 at the bag opening.

[0077] In this embodiment, the bag 40 may not be provided with the membrane 50, and the bag opening may be tightened during the stirring process by tightening the rope 41; after the stirring is completed, the bag opening of the bag 40 may be opened by pulling the bag opening.

[0078] The above are only preferred embodiments of the present application and are not intended to limit the present utility model. Although the present application has been described in detail with reference to the examples, those skilled in the art can still modify the technical solutions recorded in the aforementioned examples or make equivalent substitutions for some of the technical features therein. However, any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. An alkaline cracking device, characterized in that: include: A container (10), a stirring paddle (20) and a motor (30); The stirring paddle (20) is rotatably disposed in the container (10); The motor (30) is drivingly connected to the stirring paddle (20) and is used to drive the stirring paddle (20) to rotate; The stirring propeller (20) is an anchor propeller, a twin propeller, a single propeller or a multi-blade propeller.

2. The alkaline lysis device according to claim 1, characterized in that: The ratio of the diameter of the stirring paddle (20) to the inner diameter of the container (10) is 0.25 to 0.

5.

3. The alkaline lysis device according to claim 2, characterized in that: The stirring paddle (20) comprises an axis (214) located in the middle, a radial portion (212) extending in the radial direction, and two axial portions (211) extending in the axial direction; The axis (214), the radial portion (212) and the two axial portions (211) form an anchor-type paddle.

4. The alkaline cracking device according to claim 3, characterized in that: The ratio of the axial height of the axial portion (211) to the diameter of the stirring paddle (20) is 0.8 to 1.

2.

5. The alkaline cracking device according to claim 4, characterized in that: The radial width of the axial portion (211) is 0.1 to 0.2 times the diameter of the stirring paddle (20); The axial height of the radial portion (212) is 0.1 to 0.2 times the diameter of the stirring paddle (20); The diameter of the shaft (214) is 0.08 to 0.2 times the diameter of the stirring paddle (20).

6. The alkaline lysis device according to claim 3, characterized in that: An axial extension portion (213) is connected between the radial portion (212) and the axis (214); The axial extension portion (213) is used to increase the connection area between the radial portion (212) and the axis (214).

7. The alkaline lysis device according to claim 6, characterized in that: The axial height of the axial extension portion (213) is ≤ 1 / 3 of the height of the axial portion (211); The radial width of the axial extension portion (213) is ≤ 1 / 4 of the diameter of the stirring paddle (20).

8. The alkaline lysis device according to claim 2, characterized in that: When the stirring paddle (20) is a multi-blade paddle, the stirring paddle (20) comprises a plurality of paddle blades (21) evenly distributed around the circumference, and the paddle blades (21) are arranged obliquely.

9. The alkaline lysis device according to claim 1, characterized in that: The ratio of the height of the stirring paddle (20) along the axial direction to the diameter of the stirring paddle (20) is 0.8 to 1.

2.

10. The alkaline lysis device according to any one of claims 1 to 9, characterized in that: The ratio of the height to the inner diameter of the container (10) is 1 to 1.

5.

11. The alkaline lysis device according to any one of claims 1 to 9, characterized in that: The ratio of the height to the inner diameter of the container (10) is greater than 1.5; The stirring paddle (20) is provided with a plurality of paddle blades (21) at intervals along the axial direction.

12. The alkaline lysis device according to any one of claims 1 to 9, characterized in that: Also includes: Bagging (40); The sleeve bag (40) is sleeved in the container (10); The stirring paddle (20) extends into the bag (40).

13. The alkaline lysis device according to claim 12, characterized in that: Also includes: Diaphragm (50); The bag (40) is an open bag; The membrane (50) is placed in the bag (40) to cover the top of the bag (40); The diaphragm (50) is hung in the container (10); The stirring paddle (20) passes through the membrane (50) and extends into the bag (40).

14. The alkaline cracking device according to claim 12, characterized in that: The bag (40) is a retractable bag with a tension rope (41) disposed at the bag opening.

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