WO2016192012A1

WO2016192012A1 - Plasmid purification system

Info

Publication number: WO2016192012A1
Application number: PCT/CN2015/080509
Authority: WO
Inventors: 易初丽
Original assignee: 易初丽
Priority date: 2015-06-01
Filing date: 2015-06-01
Publication date: 2016-12-08

Abstract

A plasmid purification system, comprising: a first test tube apparatus (1) and a second test tube apparatus (2) disposed below the first test tube apparatus (1). The first test tube apparatus (1) comprises: a first test tube barrel (3), a hollow sleeve (4), and a shaft column (5), a first through hole (7) and an injection hole (7a) being provided at the top of the first test tube barrel (3), one end of the sleeve (4) being located in the first test tube barrel (3), the end, located in the first test tube barrel (3), of the sleeve (4) being provided with a stirring plate (6), the other end of the sleeve (4) extending out of the first test tube barrel (3) from the first through hole (7), and a second through hole (8) being provided at the bottom of the first test tube barrel (3). The second test tube apparatus (2) comprises: a second test tube barrel (10), a drainage tube (13), a filter apparatus located in the second test tube barrel (10), and a plasmid adsorption tube (14) located below the second test tube barrel (10) and communicated with the second test tube barrel (10).

Description

Plasmid purification system

Technical field

The invention relates to the field of experimental instruments, and in particular to a plasmid purification system.

Background technique

A plasmid is an extrachromosomal stable genetic factor. The plasmid can be artificially and intentionally added to the known or unknown DNA fragments in vitro to form recombinant "plasmid DNA", and the artificially synthesized new recombinant "plasmid DNA" can be injected into the bacteria and The bacteria multiply and multiply, then collect this large amount of bacteria, and use a special method to extract or separate the new recombinant "plasmid DNA" from the bacteria to obtain a relatively large amount (a few micrograms or several grams). There is a biologically active new recombinant "plasmid DNA". This new recombinant "plasmid DNA" can be used in a variety of scientific experiments.

The new recombinant "plasmid DNA" is extracted and purified by the kit. The extraction and purification process requires repeated centrifugation and repeated rinsing, which makes the extraction and purification process of the recombinant "plasmid DNA" time-consuming, and generally takes about 45 minutes.

Summary of the invention

In order to solve the problem of the time-consuming problem of extracting and purifying recombinant "plasmid DNA" in the prior art, the present invention provides a plasmid purification system. The technical solution is as follows:

An embodiment of the present invention provides a plasmid purification system, the plasmid purification system comprising: a first tube device and a second tube device disposed under the first tube device, the first tube device comprising: a first tube a tube, a hollow sleeve and a shaft column, the top of the first tube tube is provided with a first through hole and an injection hole, one end of the sleeve is located in the first tube tube, and the sleeve is located at the One end of the first test tube is mounted with a stirring plate, the other end of the sleeve is extended from the first through hole, and the bottom of the first test tube is provided with a second pass. a hole, the second through hole is provided with a plug matching the second through hole, one end of the shaft is connected to the plug through the sleeve, and the other end of the shaft Extending from the top of the sleeve;

The second test tube device comprises: a second test tube, a drainage tube, a filtering device located in the second test tube, and a plasmid adsorption tube located below the second test tube and in communication with the second test tube; The second test tube is located below the second through hole, the drainage tube is disposed on the outer wall of the second test tube, and the bottom end of the drainage tube extends into the plasmid adsorption tube, the plasmid A DNA adsorption layer is disposed in the adsorption tube, and a liquid outlet is disposed at a bottom of the plasmid adsorption tube.

Specifically, a bottom of the second test tube is provided with a connecting sleeve, and an outer diameter of the connecting sleeve is matched with an inner diameter of the plasmid adsorption tube, and the plasmid adsorption tube is installed through the connecting sleeve The bottom of the second test tube is provided with a through hole through which the drainage tube passes, and the bottom end of the drainage tube extends through the through hole into the plasmid adsorption tube.

Specifically, the top end of the drainage tube is provided with a cover plate.

Further, the plasmid purification system further includes: a cover driving device, the cover driving device includes: a cover motor, a cover coupling, a cover screw, a cover screw nut, a cover rail and a cover a plate pusher, the cover coupling is mounted on an output shaft of the cover motor, and the cover screw is coaxially connected to an output shaft of the cover motor through the cover coupling, The axis of the cover push rod is arranged in a horizontal direction, and the axial direction of the cover rail is the same as the axial direction of the cover screw and the axis of the cover push rod, the cover a lead screw nut is mounted on the cover screw and cooperates with the cover screw, and the cover screw nut is slidably coupled to the cover rail, and one end of the cover push rod is fixed On the cover screw nut, the other end of the cover push rod is fixed on the cover plate.

Specifically, the plasmid purification system further includes a vacuum box including: a housing, a third test tube, and a push rod, the third test tube is located inside the housing, and one end of the push rod passes through The housing is fixed on the third test tube, and a top of the housing is provided with a liquid inlet passage, and the liquid inlet passage is connected to the liquid outlet, and when the plasmid purification system receives a plasmid, the push The rod moves the third tube below the leak tube.

Further, the plasmid purification system further includes a vacuum box driving device including a first motor, a first coupling, a first lead screw, a first lead screw nut, and a first guide rail, the first a coupling is mounted on an output shaft of the first motor, the first lead screw is coaxially connected to an output shaft of the first motor through the first coupling, an axis of the first rail The direction is the same as the axial direction of the first lead screw, the first lead screw nut is mounted on the first lead screw and cooperates with the first lead screw, and the first lead screw nut and the The first rail is slidably connected, and one end of the push rod that is not connected to the third tube is fixed to the first lead screw nut.

Specifically, the plasmid purification system further includes a sleeve driving device, the sleeve driving device comprising: a second motor, a second coupling, a second lead screw, a second lead screw nut, and a second fixing a plug on the lead screw nut, the second coupling is mounted on an output shaft of the second motor, and the second lead screw passes through the output shaft of the second coupling and the second motor a coaxial connection, the second lead screw nut being mounted on the second lead screw and cooperating with the second lead screw, the axial direction of the second lead screw being the same as the axial direction of the sleeve, The sleeve is provided with a first handle, and the plug is mounted on the first handle.

Further, the plasmid purification system further includes a shaft driving device, the shaft driving device comprising: a third motor, a third coupling, a transmission shaft, an eccentric wheel, a jack and a bearing, the third coupling Mounted on an output shaft of the third motor, the drive shaft is drivingly coupled to the third coupling, the jack is fixed to the bearing, and the eccentric is mounted on the drive shaft The bearing is mounted on the ram and is drivingly connected to the eccentric. The plane of the eccentric wheel is disposed in a plane parallel to the plane of the axis of the shaft, and the bracket is provided with a pass. a hole through which the jack is drivingly coupled to the shaft, and a spring is disposed between the bracket and the bearing.

Specifically, the plasmid purification system further includes an injection device, the injection device includes: an injection device drive device and a cylinder for accommodating the reagent, the rodless cavity of the cylinder is provided with a liquid injection tube and An injection hole or a conduit through which the drainage tube communicates.

Further, the plasmid purification system includes a plurality of the cylinders and moving devices arranged side by side, the moving device including a fifth motor, a fifth lead screw mounted on the output shaft of the fifth motor, and the a fifth lead screw nut matched with a fifth lead screw and a support plate fixed to the fifth lead screw nut, wherein the injection device is fixed on the pallet, and the axial direction of the fifth lead screw is respectively The axial direction of the cylinder and the axial direction of the shaft are perpendicular to each other.

The technical solution provided by the embodiments of the present invention has the beneficial effects that the plasmid purification system provided by the embodiment of the present invention can fully mix the liquid to be extracted and the reagent by moving the sleeve, and then mix the sufficient liquid by moving the shaft column. The second through hole flows into the second test tube device, and the filter tube and the filter mesh of the second test tube device can achieve the purpose of filtering, and at the same time, the DNA adsorption layer of the second test tube device can adsorb the plasmid DNA and pass through the drainage tube to the second The eluent is added to the tube, and the eluent can be used to discharge the plasmid DNA adsorbed on the adsorption tube along with the eluent. The plasmid purification system provided in the embodiment of the invention avoids the use of the centrifuge, thereby avoiding repeated use in the centrifuge. The operation of picking and dropping saves the operation time. The extraction and purification process can be completed in a few minutes.

DRAWINGS

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly described below. It is obvious that the drawings in the following description are only some embodiments of the present invention. Other drawings may also be obtained from those of ordinary skill in the art in light of the inventive work.

1 is a schematic structural view of a first test tube device according to an embodiment of the present invention;

2 is a schematic cross-sectional structural view of a first test tube device according to an embodiment of the present invention;

3 is a schematic structural view of a second test tube device according to an embodiment of the present invention;

4 is a schematic side view showing the structure of a plasmid purification system according to an embodiment of the present invention;

Figure 5 is a schematic front view showing the structure of a plasmid purification system according to an embodiment of the present invention;

FIG. 6 is a schematic structural view of the insertion and insertion of the first handle according to the embodiment of the present invention.

detailed description

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

Example

The embodiment of the present invention provides a plasmid purification system. As shown in FIG. 1 , the plasmid purification system comprises: a first test tube device 1 comprising: a first test tube 3, a hollow sleeve 4 and a shaft The column 5, the top of the first test tube 3 is provided with a first through hole 7 (see Fig. 2) and an injection hole 7a (see Fig. 2), one end of the sleeve 4 is located in the first test tube 3, and the sleeve 4 is located One end of the first test tube 3 is mounted with a stirring plate 6 . The other end of the sleeve 4 extends from the first through hole 7 outside the first test tube 3 , and the bottom of the first test tube 3 is provided with a second through hole 8 . The second through hole 8 is provided with a plug 9 matched with the second through hole 8. One end of the shaft post 5 is connected to the plug 9 through the sleeve 4, and the other end of the shaft post 5 is extended from the top of the sleeve 4. The size of the plug 9 is matched with the size of the second through hole 8, and the axial direction of the shaft 5 can be the same as the axial direction of the sleeve 4. Therein, the injection holes 7a may be four, and the four injection holes 7a may each have a sector shape, and the four injection holes 7a may be uniformly arranged on the top of the first test tube 3.

In conjunction with FIG. 3, the plasmid purification system further includes a second test tube device 2 disposed under the first test tube device 1. The second test tube device 2 includes a second test tube 10, a drainage tube 13, and a second test tube 10. a filter device and a plasmid adsorption tube 14 located below the second test tube 10 and communicating with the second test tube 10, the second test tube 10 is located below the second through hole 8, and the drainage tube 13 is disposed on the outer wall of the second test tube 10. The bottom end of the drainage tube 13 extends into the plasmid adsorption tube 14, and the DNA adsorption layer 14a is disposed in the plasmid adsorption tube 14, and the liquid outlet 21 is provided at the bottom of the plasmid adsorption tube 14. The filter device may include a filter tube 11 and a filter screen 15, which may be disposed in the second test tube 10, the filter screen 15 may be disposed on the bottom inner wall of the second test tube 10, and the filter tube 11 is located in the first test tube device Between 1 and the filter 15, the filter 15 provided in this embodiment is denser than the filter on the filter tube 11, the filter 11 can filter larger impurities, and the filter 15 can be further filtered. The impurity, the DNA adsorption layer 14a is used to adsorb the plasmid, and the DNA adsorption layer 14a may be disposed on the bottom inner wall of the plasmid adsorption tube 14.

Wherein, the top of the filter tube 11 may be provided with an outwardly extending outer edge, and the inner wall of the second test tube 10 may be provided with a protrusion for arranging the outer edge of the filter tube 11, and the filter tube 11 can be placed on the convex portion through the outer edge thereof. Start up. Further, the first test tube device 1 and the second test tube device 2 may be arranged in a plurality of side by side, and the first test tube device 1 is disposed above the second test tube device 2, and the number of the first test tube device 1 and the second test tube device 2 may be The number of samples actually needed to be inspected is adjusted.

Specifically, a liquid outlet pipe 10a may be disposed on the bottom outer wall of the second test tube tube 10, and the second test tube tube 10 communicates with the plasmid adsorption tube 14 through the liquid discharge pipe 10a.

Specifically, the bottom of the second test tube 10 may be provided with a connecting sleeve 10b. The outer diameter of the connecting sleeve 10b is matched with the inner diameter of the plasmid adsorption tube 14, and the plasmid adsorption tube 14 is mounted on the second test tube 10 through the connecting sleeve 10b. At the bottom of the connecting sleeve 10b, a through hole for the passage of the drainage tube 13 is opened, and the bottom end of the drainage tube 13 extends through the through hole into the plasmid adsorption tube 14. Wherein, the connecting sleeve 10b can be a cylindrical structure, and the connecting sleeve 10b can be arranged around the liquid outlet tube 10a, and the connecting sleeve 10b can be integrated with the second test tube 10. In addition, the connecting sleeve 10b can be movably mounted together with the plasmid adsorption tube 14, that is, the connecting sleeve 10b is stuck on the inner wall of the plasmid adsorption tube 14, or the external wall of the connecting sleeve 10b can be provided with an external thread on the inner wall of the plasmid adsorption tube 14. The internal thread is arranged, and the connecting sleeve 10b and the plasmid adsorption tube 14 can be screwed together, and the movable sleeve 10b and the plasmid adsorption tube 14 can facilitate the removal of the plasmid adsorption tube 14 to facilitate the replacement of the DNA inside the plasmid adsorption tube 14. Adsorption layer 14a.

Specifically, the top end of the draft tube 13 may be provided with a cover plate 13a. The top end of the draft tube 13 may be flush with the top of the second test tube 10, and the drain tube 13 is disposed on the outer wall of the second test tube 10, and when the second test tube 10 is processed, the wall of the second test tube 10 The drainage tube 13 is machined such that the second tube 10 and the drainage tube 13 are integrated. The cover plate 13a is provided to prevent foreign matter from falling into the drainage tube 13.

Specifically, as shown in FIG. 4, the plasmid purification system may further include: a cover driving device. In this embodiment, the cover driving device comprises: a cover motor 64, a cover coupling 65, a cover screw 66, a cover screw nut 67, a cover rail 68 and a cover push rod 69, and a cover The plate coupling 65 is mounted on the output shaft of the cover motor 64, and the cover screw 66 is coaxially connected to the output shaft of the cover motor 64 through the cover coupling 65, and the axial direction of the cover push rod 69 is horizontal The direction is arranged, and the axial direction of the cover rail 68 is the same as the axial direction of the cover screw 66 and the axial direction of the cover push rod 69, and the cover screw nut 67 is mounted on the cover screw 66 and the cover The lead screw 66 is matched, and the cover screw nut 67 is slidably connected to the cover rail 68. One end of the cover push rod 69 is fixed on the cover screw nut 67, and the other end of the cover push rod 69 is fixed to the cover plate. 13a. It will be readily appreciated that in other embodiments, the cover drive can be manually operated by manually pushing and pulling the cover pusher 69 to effect horizontal movement of the cover 13a.

The drainage tube 13 can be two, symmetrically arranged on the outer wall of the second test tube 10, and the cover plate 13a is also arranged correspondingly. The connection end of the cover push rod 69 and the cover plate 13a can be U-shaped, and the connection end is U. The type of cover pusher 69 can push the two cover plates 13a at the same time.

Specifically, as shown in FIG. 4, the plasmid purification system may further include a vacuum box 16 including: a housing 17, a third tube 18, and a push rod 26, the third tube 18 being located inside the housing 17, and the push rod 26 One end of the housing 17 is fixed to the third tube 18, and the top of the housing 17 is provided with a liquid inlet passage 19, and the liquid inlet passage 19 communicates with the liquid outlet 21, and when the plasmid purification system receives the plasmid, the push rod 26 will The third test tube 18 is moved below the liquid outlet 21 . When the plasmid purification system does not collect a plasmid (such as a waste liquid), the third tube 18 can be moved to a position other than below the liquid outlet 21. The vacuum box 16 can provide a negative pressure, and when the second tube device 2 discharges liquid, the plasmid and the waste liquid in the second tube device 2 can be extracted, and the residual material on the tube wall can be reduced to ensure the second tube device. 2 The discharged plasmid is relatively pure.

Further, as shown in FIG. 4, in the embodiment, the plasmid purification system may further include a vacuum box driving device, and the vacuum box driving device may include a first motor 22, a first coupling 23, and a first lead screw 24. a first lead screw nut 25 and a first guide rail 27, the first coupling 23 is mounted on the output shaft of the first motor 22, and the first lead screw 24 passes through the first coupling 23 and the output shaft of the first motor 22. Coaxially connected, the axial direction of the first guide rail 27 is the same as the axial direction of the first lead screw 24, and the first lead screw nut 25 is mounted on the first lead screw 24 and cooperates with the first lead screw 24, the first lead screw The nut 25 is slidably coupled to the first rail 27, and one end of the push rod 26 that is not connected to the third tube is fixed to the first lead screw nut 25. It is easy to know that in other embodiments, the vacuum box 16 can also push and pull the push rod 26 in a manual manner to achieve the purpose of pushing and pulling the third tube 18. The first guide rail 27 can ensure the movement of the first lead screw nut 25 The direction is a straight line, and the operation of the first screw nut 25 can be made smoother by mounting the first rail 27. Further, a slider may be provided on the first lead screw nut 25 to mount the slider on the first guide rail 27, and the operation of the first lead screw nut 25 may be made smoother by mounting the slider.

Specifically, the second test tube device 2 may further include a leak tube 20 disposed on the liquid outlet 21, and the leak tube 20 is in communication with the liquid outlet 21, and the inner wall of the liquid inlet passage 19 is provided with an internal thread and a drain The outer wall of the tube 20 is provided with an external thread that cooperates with the internal thread on the inlet passage 19, and the second tube device 2 is mounted on the inlet passage 19 by internal threads, and the inner wall of the leakage tube 20 is of a funnel type.

Further, as shown in FIG. 5, the top of the side wall of the vacuum box 16 may be provided with a first ball valve 56 for controlling the degree of vacuum of the vacuum box 16, and the bottom portion of the side wall of the vacuum box 16 may also be arranged for discharging waste. The second ball valve 57 of the liquid. The inside of the vacuum box 16 may be provided with a water tank, and the water tank communicates with the second ball valve 57.

Specifically, as shown in FIG. 1 and FIG. 4, the plasmid purification system may further include a support 28, and the support 28 may be provided with a test tube support plate 29 and a test tube lower plate 30, and the first test tube tube 3 is respectively provided with a groove 31 and the funnel-shaped outer edge, the test tube upper plate 29 and the test tube lower plate 30 are respectively provided with holes matching the groove 31 and the outer edge, and the first test tube 3 is mounted on the test tube through the groove 31 and the outer edge. Upper plate 29 and test tube lower plate 30. Wherein, the side wall of the test tube 29 and the lower tube 30 of the test tube may be provided with a limited position protrusion, and the support 28 may also be provided with a mounting groove, and the size of the installation groove is respectively matched with the upper plate 29 and the test tube The size of the plate 30 is matched, and the side wall of the mounting groove can be provided with a limiting slot matched with the limiting protrusion. The cooperation of the limiting protrusion and the limiting slot can ensure the stable installation of the first test tube device 1 On the bracket 28. When the plasmid purification system is assembled, the first test tube device 1 can be first mounted on the test tube support plate 29 and the test tube lower plate 30, and the test tube upper plate 29 and the test tube lower plate 30 can be attached to the support 28.

Specifically, as shown in FIG. 4 and FIG. 6 , in the embodiment, the plasmid purification system may further include a sleeve driving device, and the sleeve driving device may include: a second motor 39, a second coupling 40, and a a second lead screw 41, a second lead screw nut 42 and a plug 43 fixed to the second lead screw nut 42, the second coupling 40 is mounted on the output shaft of the second motor 39, and the second lead screw 41 is passed through The second coupling 40 is coaxially connected to the output shaft of the second motor 39, and the second spindle nut 42 is mounted on the second lead screw 41 and cooperates with the second lead screw 41, and the axial direction of the second lead screw 41 is The sleeve 4 has the same axial direction, and the sleeve 4 may be provided with a first handle 44, and the insertion 43 is mounted on the first handle 44. It will be readily appreciated that in other embodiments, the sleeve drive can be manual, i.e., the sleeve 4 can also be moved up and down manually. among them, The plug 43 is fixed to one end of the second lead screw nut 42 and may be provided with a socket, and the first handle 44 may be disposed in the socket.

Further, as shown in FIG. 5, in this embodiment, the plasmid purification system may further include a shaft driving device, and the shaft driving device may include: a third motor 32, a third coupling 33, a transmission shaft 34, The eccentric 35, the ram 37 and the bearing 36, the third coupling 33 is mounted on the output shaft of the third motor 32, the transmission shaft 34 is drivingly coupled to the third coupling 33, and the ram 37 is fixed to the shaft 5 The eccentric 35 is mounted on the drive shaft 34. The bearing 36 is mounted on the jack 37 and is drivingly coupled to the eccentric 35. The plane in which the direction of rotation of the eccentric 35 lies is parallel to the plane of the axis of the shaft 5, on the bracket 28. A through hole is provided, and the jack 37 is drivingly connected to the shaft post 5 through the through hole, and a spring 38 is disposed between the bracket 28 and the bearing 36. It is easy to know that in other embodiments, the driving mode of the shaft column 5 can be manual, that is, the shaft column 5 can also be moved up and down by manual means. The ram 66 and the shaft post 5 can be connected by a flange 36a. Further, a second handle can be mounted on the shaft post 5, and the ram 37 is connected to the shaft post 5 via the second handle. During the movement of the eccentric 35, the shaft column 5 can be pressed by the ejector lever 37 to achieve the downward movement of the shaft column 5, and the spring 38 is pressed. With the rotation of the eccentric wheel 35, the eccentric wheel 35 is no longer squeezed. When the shaft column 5 is pressed, the spring 38 can help the shaft column 5 to be reset. Furthermore, two lifting lugs can be provided on the jack 37 for mounting the bearing 36. In this embodiment, the first test tube device 1 can be arranged six, then the number of the eccentric 35, the ram 37 and the bearing 36 is evenly the same as the number of the first test tube device 1, that is, six, and the eccentric wheel The plane in which the direction of rotation of 35 is located may coincide with the plane of the axis of the shaft column 5.

In the embodiment, the plug-in slider 58 can also be mounted on the plug-in strip 43. The bracket 28 is mounted with a slide rail 59 that cooperates with the plug-in slider 58, and the axial direction of the slide rail 59 and the second wire The axes of the bars 41 are arranged in parallel.

Specifically, the plasmid purification system further includes an injection device, which may include a cylinder 49 for accommodating the reagent and an injection device driving device, and the rodless chamber of the cylinder 49 is provided with a liquid injection tube 50 and the injection hole 7a or drainage A conduit 51 in which the tube 13 is in communication. The check valve 50 and the conduit 51 may be respectively disposed with a check valve 50a for preventing backflow of the reagent in the cylinder 49. Further, the duct 51 communicating with the injection hole 7a and the duct 51 communicating with the draft tube 13 may be disposed on different cylinders 49.

Specifically, as shown in FIG. 4, in the embodiment, the injection device driving device includes: a fourth motor 45, a fourth coupling 46, a fourth lead screw 47, a fourth lead screw nut 48, and an injection device guide rail. 12, the fourth coupling 46 is mounted on the output shaft of the fourth motor 45, the fourth lead screw 47 is coaxially connected to the output shaft of the fourth motor 45 through the fourth coupling 46, and the fourth lead screw nut 48 is mounted. On the fourth lead screw 47 and The fourth lead screw 47 is engaged, the fourth lead screw nut 48 is coupled to the piston rod of the cylinder 49, and the fourth lead screw nut 48 is mounted on the injection device guide 12, the axis of the injection device guide 12 and the fourth lead screw 47. The axes are arranged in parallel. Wherein, the fourth lead screw nut 48 is in driving contact with the piston rod of the cylinder 49, that is, the fourth lead screw nut 48 can push the piston rod of the cylinder 49 to move toward the rodless cavity of the cylinder 49, thereby realizing the upcoming cylinder The purpose of the reagent extrusion in 49. It will be readily appreciated that in other embodiments, the manner of driving the injection device can be manual.

Specifically, the injection device may further include two support plates 63. The two ends of the injection device guide rail 12 are respectively fixed on the two support plates 63, and the two support plates 63 may be disposed on the bracket 28.

Further, as shown in FIG. 4 and FIG. 5, the plasmid purification system may include a plurality of cylinders 49 and moving devices arranged side by side, and the moving device includes a fifth motor 52 and a fifth wire mounted on the output shaft of the fifth motor 52. a bar 53, a fifth lead screw nut 54 that cooperates with the fifth lead screw 53, and a bracket 55 fixed to the fifth lead screw nut 54, the injection device is fixed to the pallet 55, and the axial direction of the fifth lead screw 53 The axial direction of the cylinder 49 and the axial direction of the shaft 5 are perpendicular to each other, respectively. Among them, a plurality of cylinders 49 arranged side by side are used to accommodate a plurality of reagents to prevent mutual contamination of the reagents. Wherein, the cylinder 49 may be plural, and the plurality of cylinders 49 may respectively accommodate the solution P1, the solution P2, the solution P3, the solution P4, the isopropanol, the equilibrium liquid BL, the deproteinized liquid PD, the rinse liquid PW and the elution buffer. Liquid TB.

Further, the cover motor 64, the second motor 39, the third motor 32, and the fourth motor 45 may be fixed to the bracket 28. In the present embodiment, the pallet 55 may be disposed on the horizontal plate 60 of the bracket 28, and the horizontal plate 60 and the pallet 55 may be provided with guiding means. As shown in FIG. 4, the guiding device may further include a second guide 61. And the second slider 62, the moving device may be disposed in the middle of the horizontal plate 60 and the pallet 55, and the guiding device may be two, disposed on both sides of the horizontal plate 60 and the pallet 55, and the second of the two guiding devices The axes of the guide rails 61 are each arranged in parallel with the axis of the fifth lead screw 53 of the moving device. In addition, the two guides ensure that the pallet 55 runs smoothly.

The process of extracting a plasmid by using the plasmid purification system provided by the embodiment of the present invention is as follows:

The bacterial suspension to be extracted is placed in the first test tube device 1, as shown in FIG. 2, by manually or driving the fifth motor 52, the transfer plate 55 is moved to move the fifth screw nut 54 to the desired reagent. At the piston rod of the cylinder 49, as shown in FIG. 4, the fourth motor 45 is manually or driven to move the fourth screw nut 48 along the fourth lead screw 47, and the reagent in the cylinder 49 is injected through the injection hole 7a. In a test tube device 1, the sleeve 4 is moved up and down to achieve mixing, and at the same time, the agitating plate 6 on the sleeve 4 enables the bacteria and the reagent to be sufficiently mixed. If there are multiple samples, you can continue to move the pallet 55 and add it to the next first tube device 1. Enter the required reagents. Pressing the shaft column 5 downwards causes the plug 9 to be disengaged from the second through hole 8. The liquid in the first test tube device 1 flows through the second through hole 8 to the filter tube 11 of the second test tube device 2 and the filter screen 15 in sequence. Bulk impurities can be filtered through the filter tube 11, while minute impurities are filtered through the filter 15. At this time, the plasmid and the reagent in the extract are passed through the filter tube 11 and the filter 15 to the plasmid adsorption tube 14, and the plasmid is adsorbed onto the DNA adsorption layer 14a, and the waste liquid can be discharged from the plasmid adsorption tube 14 and Draw into the vacuum box under the negative pressure of the vacuum box. Manually or driving the cover motor 64, the cover screw nut 67 moves to push the cover pusher 69 to move the cover 13a away from the drain tube 13, so that the drain tube 13 is exposed. At this time, by manually or driving the fifth motor 52, the moving plate 55 moves the fifth screw nut 54 to the piston rod of the cylinder 49 containing the rinsing liquid, and manually or drives the fourth motor 45 to make the fourth wire. The lever nut 48 moves along the fourth lead screw 47, and the reagent in the cylinder 49 is injected into the drainage tube 13 through the conduit 51 and flows into the plasmid adsorption tube 14, and the waste liquid can flow out through the plasmid adsorption tube 14 and in the vacuum box. The vacuum is sucked into the vacuum box, and by manually or driving the fifth motor 52, the moving plate 55 moves the fifth screw nut 54 to the piston rod of the cylinder 49 containing the eluent, manually or driven. The fourth motor 45 moves the fourth lead screw nut 48 along the fourth lead screw 47, and the reagent in the cylinder 49 is injected into the drainage tube 13 through the conduit 51 and flows into the plasmid adsorption tube 14, by moving the push rod 26, The third tube 18 is moved below the liquid outlet 21, and the plasmid is drawn into the third tube 18 by evacuation to obtain an extracted plasmid.

The plasmid purification system provided by the embodiment of the invention can fully mix the liquid to be extracted and the reagent by moving the sleeve, and then flow the mixed liquid from the second through hole to the second test tube device through the moving shaft column, and then pass through the second The filter tube of the test tube device and the filter mesh can achieve the purpose of filtration. At the same time, the DNA adsorption layer of the second test tube device can adsorb the plasmid DNA, and the eluent can be added to the second test tube through the drainage tube, and the eluent can be adsorbed on the eluate. The plasmid DNA on the adsorption tube is discharged together with the eluent. The plasmid purification system provided in the embodiment of the invention avoids the use of the centrifuge, thereby avoiding repeated operations in the centrifuge, saving operation time, and the entire extraction and purification process. Can be completed in a few minutes.

The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalents, improvements, etc., which are within the spirit and scope of the present invention, should be included in the protection of the present invention. Within the scope.

Claims

A plasmid purification system, characterized in that the plasmid purification system comprises: a first tube device and a second tube device disposed under the first tube device,

The first test tube device comprises: a first test tube, a hollow sleeve and a shaft column, the top of the first test tube is provided with a first through hole and an injection hole, and one end of the sleeve is located at the first In the test tube, an end of the sleeve located in the first test tube is mounted with a stirring plate, and the other end of the sleeve is protruded from the first through hole outside the first test tube. a second through hole is disposed in a bottom portion of the first test tube, and a plug matching the second through hole is disposed in the second through hole, and one end of the shaft passes through the sleeve and the a plug connection, the other end of the shaft is extended by the top of the sleeve;

The second test tube device comprises: a second test tube, a drainage tube, a filtering device located in the second test tube, and a plasmid adsorption tube located below the second test tube and in communication with the second test tube; The second test tube is located below the second through hole, the drainage tube is disposed on the outer wall of the second test tube, and the bottom end of the drainage tube extends into the plasmid adsorption tube, the plasmid A DNA adsorption layer is disposed in the adsorption tube, and a liquid outlet is disposed at a bottom of the plasmid adsorption tube.
The plasmid purification system according to claim 1, wherein a bottom portion of the second test tube is provided with a connecting sleeve, and an outer diameter of the connecting sleeve is matched with an inner diameter of the plasmid adsorption tube, a plasmid adsorption tube is installed at the bottom of the second test tube through the connecting sleeve, and the connecting sleeve is provided with a through hole through which the drainage tube passes, and the bottom end of the drainage tube passes through the through hole Extend into the plasmid adsorption tube.
The plasmid purification system according to claim 1, wherein a top end of the drainage tube is provided with a cover.
The plasmid purification system according to claim 3, wherein the plasmid purification system further comprises: a cover driving device, wherein the cover driving device comprises: a cover motor, a cover coupling, and a cover screw a cover screw nut, a cover rail and a cover push rod, the cover coupling is mounted on an output shaft of the cover motor, and the cover screw passes through the cover coupling An output shaft of the cover motor is coaxially connected, an axis of the cover push rod is arranged in a horizontal direction, and an axial direction of the cover rail and an axial direction of the cover screw and the cover are pushed The axes of the rods are all in the same direction, the cover screw nut is mounted on the cover screw and cooperates with the cover screw, and the cover screw nut slides with the cover rail Connecting, one end of the cover push rod is fixed on the cover screw nut, The other end of the cover push rod is fixed to the cover.
The plasmid purification system according to claim 1, wherein the plasmid purification system further comprises a vacuum box, the vacuum box comprising: a housing, a third test tube, and a push rod, the third test tube being located in the shell Inside the body, one end of the push rod is fixed on the third test tube through the casing, and a top of the casing is provided with a liquid inlet passage, and the liquid inlet passage is connected with the liquid outlet, when When the plasmid purification system receives the plasmid, the push rod moves the third tube below the leak tube.
The plasmid purification system according to claim 5, wherein the plasmid purification system further comprises a vacuum box driving device, the vacuum box driving device comprising a first motor, a first coupling, a first lead screw, and a first a lead screw nut and a first guide rail, the first coupling is mounted on an output shaft of the first motor, and the first lead screw passes through an output of the first coupling and the first motor The shaft is coaxially connected, the axial direction of the first guide rail is the same as the axial direction of the first lead screw, and the first lead screw nut is mounted on the first lead screw and is opposite to the first lead screw The first lead screw nut is slidably coupled to the first guide rail, and one end of the push rod that is not connected to the third test tube is fixed to the first lead screw nut.
The plasmid purification system according to claim 1, wherein the plasmid purification system further comprises a sleeve driving device, the sleeve driving device comprising: a second motor, a second coupling, a second lead screw, a second lead screw nut and a plug fixed to the second lead screw nut, the second coupling is mounted on an output shaft of the second motor, and the second lead screw passes the second a coupling coaxially connected to an output shaft of the second motor, the second lead screw nut being mounted on the second lead screw and cooperating with the second lead screw, the second lead screw The axial direction is the same as the axial direction of the sleeve, and the sleeve is provided with a first handle, and the plug is mounted on the first handle.
The plasmid purification system according to claim 1, wherein the plasmid purification system further comprises a shaft driving device, the shaft driving device comprising: a third motor, a third coupling, a transmission shaft, and an eccentric wheel a ram and a bearing, the third coupling is mounted on an output shaft of the third motor, the transmission shaft is drivingly coupled to the third coupling, and the ram is fixed on the bearing The eccentric is mounted on the transmission shaft, the bearing is mounted on the ram and is drivingly connected with the eccentric, the plane of rotation of the eccentric is on a plane with the axis of the shaft The planes are arranged in parallel, and the bracket is provided with a through hole, and the jack is drivenly connected to the shaft through the through hole, and a spring is arranged between the bracket and the bearing.
The plasmid purification system according to claim 1, wherein said plasmid purification system An injection device is further included, the injection device includes: an injection device driving device and a cylinder for accommodating the reagent, the rodless cavity of the cylinder is provided with a liquid injection tube and is connected to the injection hole or the drainage tube Catheter.
The plasmid purification system according to claim 9, wherein said plasmid purification system comprises a plurality of said cylinders and moving devices arranged side by side, said moving means comprising a fifth motor mounted on said fifth motor a fifth lead screw on the output shaft, a fifth lead screw nut mated with the fifth lead screw, and a pallet fixed to the fifth lead screw nut, the injection device being fixed on the pallet The axial direction of the fifth lead screw is perpendicular to the axial direction of the cylinder barrel and the axial direction of the shaft column, respectively.