Plasmid extraction column passing device

Abstract

The utility model discloses a plasmid extraction column passing device which comprises a controller with a cooling unit, multi-section type telescopic rods on the controller are connected with a pressing plate, and a plurality of cooling grooves are arranged between the two multi-section type telescopic rods; a first layer plate and a second layer plate are stacked between the pressing plate and the controller, a plurality of first column passing channels with built-in filter columns are uniformly distributed on the first layer plate, a plurality of second column passing channels with built-in adsorption columns are uniformly distributed on the second layer plate, the first column passing channels and the second column passing channels are consistent in size, a first conical liquid outlet is formed in the lower end of each first column passing channel, and a second conical liquid outlet is formed in the lower end of each second column passing channel. A second conical liquid outlet is formed in the lower end of the second column passing channel; sample adding pipes in one-to-one correspondence with the first column passing channels are arranged on the pressing plate, a high-pressure air pump is arranged in the controller, an air blowing pipe is arranged on the high-pressure air pump, and air outlet branch pipes communicated with the sample adding pipes one by one are arranged at the air outlet end of the air blowing pipe. The utility model has the advantages that the plasmid extraction lysate can be easily filtered and adsorbed, the operation time is greatly shortened, and the plasmid structure is prevented from being damaged.

Description

Technical Field

[0001] This utility model relates to the field of plasmid extraction technology, and in particular to a plasmid extraction column device. Background Technology

[0002] Plasmids are DNA molecules other than chromosomes (or nucleoids) in organisms such as bacteria, yeast, and actinomycetes. They exist in the cytoplasm (except for yeast, where 2μm plasmids are located in the nucleus). They have the ability to replicate autonomously and express the genetic information they carry. The genetic information carried by plasmids can confer certain biological traits to the host bacteria, which is beneficial for the bacteria to survive under specific environmental conditions.

[0003] A vector is a tool used to amplify a useful target DNA fragment into a recipient cell through recombinant DNA technology. Plasmids are commonly used vectors in recombinant DNA technology. They utilize their self-replication function to amplify the genetic information they carry after entering the cell. This plasmid transfection operation is widely used in basic experiments.

[0004] The traditional plasmid extraction process is as follows: First, E. coli colonies containing the target plasmid are inoculated into liquid culture medium and cultured overnight with constant temperature and shaking to complete the bacterial culture. Second, the cultured bacterial solution is centrifuged in an EP tube and the supernatant is removed to complete the bacterial enrichment. Third, lysis buffer is added to the precipitated bacterial solution to release plasmid DNA and complete cell lysis. Fourth, the supernatant of the lysed bacterial solution is added to a syringe-type collection tube with a filter column for filtration to remove large molecular impurities. The filtrate is then added to a syringe-type collection tube with an adsorption column for adsorption. After adsorption, the adsorption column is rinsed twice. After rinsing, elution buffer is added to the adsorption column to fully elute the plasmid DNA, and the eluent containing purified plasmid DNA is collected to complete the entire plasmid extraction process.

[0005] In the above plasmid extraction process, since the liquid flow rate through the filter column and adsorption column is relatively slow, high-speed centrifugation is usually required to accelerate the liquid flow rate. High-speed centrifugation may cause damage to the plasmid structure. In addition, the repeated loading and unloading of the collection tube with the filter column or adsorption column into and out of the centrifuge is also very cumbersome, which greatly affects the experimental efficiency of plasmid extraction. Furthermore, it is necessary to maintain a low temperature environment throughout the entire plasmid extraction process to increase the stability of the plasmid. Summary of the Invention

[0006] The purpose of this invention is to address the shortcomings of existing technologies by providing a plasmid extraction column pass device that accelerates the plasmid extraction column pass process by pressurizing with an air pump.

[0007] To achieve the above objectives, the present invention can adopt the following technical solution:

[0008] The plasmid extraction column chromatography device of this invention includes a controller with a cooling unit. A pressure plate is connected to the controller via a pair of vertically arranged multi-segment telescopic rods. Multiple cooling grooves for inserting EP tubes are evenly spaced on the controller between the two multi-segment telescopic rods. A first plate and a second plate are stacked between the pressure plate and the controller, directly above the cooling grooves. The first plate has multiple first column chromatography channels with built-in filter columns, and the second plate has multiple second column chromatography channels with built-in adsorption columns. The first and second column passages have the same dimensions. The lower end of the first column passage is a first conical liquid outlet that is inserted into the second column passage, and the lower end of the second column passage is a second conical liquid outlet that is inserted into the cooling tank. The pressure plate is provided with sample tubes that correspond one-to-one with the first column passages. The controller is provided with a high-pressure gas pump for blowing nitrogen. The high-pressure gas pump is provided with a blowing pipe that extends out of the controller. The outlet end of the blowing pipe is provided with an outlet branch pipe that corresponds one-to-one with the sample tube.

[0009] Furthermore, to ensure that the high-pressure gas pump can blow high-pressure nitrogen into the first column passage as completely as possible, multiple sealing rings are evenly distributed on the bottom surface of the pressure plate, corresponding to the upper end of each of the first column passages. At this time, the sample tube is sealed and inserted into the pressure plate inside each of the sealing rings, ensuring that the gas can be completely blown into the first column passage.

[0010] Furthermore, electromagnets are provided on the left and right sides of the controller, and adsorption magnets adapted to the electromagnets are provided on the left and right sides of the bottom surface of the pressure plate. The height of the adsorption magnets should be less than or equal to the height of the first through-column channel, so that even when only one first or second layer plate is placed between the pressure plate and the controller, the electromagnets can be energized to attract the adsorption magnets downwards, and the suction force is used to increase the pressure when the pressure plate covers the upper end of the first through-column channel or the upper end of the second through-column channel.

[0011] Furthermore, for ease of operation, a touch screen timer, an air pump switch, and an electromagnetic switch should also be installed on the front wall of the controller.

[0012] Furthermore, a lifting handle is provided on the top wall of the pressure plate to facilitate manual lifting of the pressure plate after the electromagnet is de-energized.

[0013] Furthermore, each of the sample dispensing tubes is equipped with an on / off valve. In this case, the air outlet branch pipe should be connected to the sample dispensing tube on the outlet side of the on / off valve. This facilitates manual operation of the sample dispensing tube to add or stop the sample dispensing, while also ensuring that the air blowing pipe does not blow air into the liquid inlet end of the sample dispensing tube.

[0014] The advantages of this invention are that, through the first column passage on the first plate and the second column passage on the second plate, the filtration and adsorption operations of the plasmid extraction lysate can be easily achieved. A high-pressure air pump can be used to blow nitrogen into either the first or second column passage, thereby accelerating the liquid's passage through the filter or adsorption column by pressurizing with high-pressure nitrogen. This ensures no liquid contamination while significantly increasing the liquid flow rate during plasmid extraction. Compared to traditional centrifugation, this method greatly shortens the operation time, avoids plasmid structure damage, and ensures the integrity and function of the final extracted plasmid remain unaffected, improving the success rate and reliability of subsequent experiments. Furthermore, the final liquid collected in the EP tube within the cooling tank after plasmid extraction allows for continuous cooling, effectively increasing plasmid stability and further guaranteeing the quality and activity of the extracted plasmid. In addition, the entire device simplifies the cumbersome operation of repeatedly loading and unloading the centrifuge in traditional methods, reducing experimental steps, lowering operational difficulty, and improving experimental efficiency, making it suitable for large-scale plasmid extraction experiments. Attached Figure Description

[0015] Figure 1 This is a schematic diagram of the structure of this utility model.

[0016] Figure 2 yes Figure 1 A schematic diagram of the insertion of the first and second layers of the plate.

[0017] Figure 3 yes Figure 2 A cross-sectional view of the first layer of the plate.

[0018] Figure 4 yes Figure 2 A cross-sectional view of the second layer plate. Detailed Implementation

[0019] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0020] like Figure 1-4 As shown, the plasmid extraction column device of this utility model includes a controller 1. A pressure plate 3 is connected to the controller 1 by a pair of vertically arranged multi-segment telescopic rods 2. The bottom of the controller 1 has a cooling unit 4. Multiple cooling grooves 5 for fitting inserted EP tubes are evenly spaced on the top wall of the controller 1 between the two multi-segment telescopic rods 2. The cooling unit 4 can cool the EP tube inserted in the cooling groove 5, so that the liquid in the EP tube is continuously kept in a low temperature environment.

[0021] In the space between the pressure plate 3 and the controller 1, there are stacked first plates 6 and second plates 7, which are directly above the cooling tank 5. The first plate 6 has multiple first column passage channels 9 with built-in filter columns 8, and the second plate 7 has multiple second column passage channels 11 with built-in adsorption columns 10. The first column passage channels 9 and the second column passage channels 11 correspond one-to-one and have the same size. The lower end of the first column passage channel 9 is a first conical liquid outlet 12 that is inserted into the second column passage channel 11, and the lower end of the second column passage channel 11 is a second conical liquid outlet 13 that is inserted into the cooling tank 5. Thus, the first plate 6 and the second plate 7 are stacked one on top of the other and finally inserted into the cooling tank 5.

[0022] The pressure plate 3 is equipped with a sample tube 14 corresponding to the first column passage 9. The controller 1 is equipped with a high-pressure air pump 15 for blowing nitrogen. The high-pressure air pump 15 has an air blowing pipe 16 extending out of the controller 1. The air outlet of the air blowing pipe 16 is equipped with multiple air outlet branches 17 that are connected to the sample tubes 14. In order to facilitate independent control of each sample tube 14 for sample addition, an on / off valve 18 should be installed on each sample tube 14. At this time, the air outlet branches 17 should be connected to the sample tubes 14 on the outlet side of the on / off valve 18 to ensure that each sample tube 14 can be manually controlled for sample addition while ensuring that the air blowing pipe 16 does not blow air into the liquid inlet of the sample tube 14.

[0023] Furthermore, to ensure that the high-pressure gas pump 15 can blow high-pressure nitrogen into the first column passage 9 as completely as possible, multiple sealing ring gaskets 19, each corresponding to one of the first column passage 9, should be evenly distributed on the bottom surface of the pressure plate 3. The sealing ring gaskets 19 are rubber or silicone gaskets, and can be correspondingly fastened to the upper end of each first column passage 9, thus keeping the upper end of the first column passage 9 sealed. Simultaneously, the sample dispensing tube 14 should be correspondingly and sealingly inserted into the pressure plate 3 inside each sealing ring gasket 19, ensuring that the gas can be completely blown into the first column passage 9. Of course, in actual use, since the first column passage 9 and the second column passage 11 are the same size, when the first layer plate 6 is removed, the sealing ring gaskets 19 can also correspondingly fasten to the upper end of each second column passage 11.

[0024] In addition, a pair of electromagnets 20 are provided on the left and right sides of the controller 1, and adsorption magnets 21 adapted to the electromagnets 20 are provided on the left and right sides of the bottom surface of the pressing plate 3; the height of the adsorption magnet 21 should be less than or equal to the height of the first column passing channel 9. Of course, since the sizes of the first column passing channel 9 and the second column passing channel 11 are the same, the adsorption magnet 21 will also be less than or equal to the height of the second column passing channel 11, ensuring that when only one layer of the first layer plate 6 or the second layer plate 7 is placed between the pressing plate 3 and the controller 1, the electromagnet 20 can be powered on to attract the adsorption magnet 21 downward, so as to use the attraction force to increase the pressure when the pressing plate 3 presses on the upper end of the first column passing channel 9 or the upper end of the second column passing channel 11. To facilitate the control of the entire plasmid extraction column device, a touch screen timer 22, an air pump switch 23 and an electromagnetic switch 24 should also be provided on the front wall of the controller 1, and a pair of lifting handles 25 should be provided on the top wall of the pressing plate 3 to facilitate manually holding the lifting handles 25 to lift the pressing plate 3 upward after the electromagnet 20 is powered off.

[0025] When actually performing the plasmid extraction experiment: First, keep the electromagnetic switch 24 closed and lift the pressing plate 3 upward, so as to accurately insert the stacked first layer plate 6 and the second layer plate 7 into the corresponding cooling grooves 5 one by one; then release the pressing plate 3 to let it fall naturally, and make the sealing ring pads 19 on the bottom surface of the pressing plate 3 seal on each first column passing channel 9 one by one. At this time, turn on the electromagnet 20 and use the attraction force to continuously attract the adsorption magnet 21 vertically downward, so as to provide further assistance for the downward pressure of the pressing plate 3; then the plasmid-containing bacterial liquid after cell lysis can be added into each first column passing channel 9 through the sample addition tube 14, and high-pressure nitrogen can be blown into the first column passing channel 9 through the high-pressure air pump 15 in the controller 1, as well as the blowing tube 16 and the outlet branch pipe 17 matching the high-pressure air pump 15. Since nitrogen is an inert gas, it will not react with the bacterial liquid in the first column passing channel 9, and can only provide a continuous downward pressure on the bacterial liquid added into the first column passing channel 9, and the bacterial liquid can flow downward quickly through the pressure to ensure that the bacterial liquid filters impurities through the filter column 8 in the first column passing channel 9. In the plasmid extraction experiment, the main function of the filter column 8 is to remove impurities such as cell debris and proteins to ensure that the sample is relatively pure before entering the adsorption column 10. Its pore size is usually about 20 - 30μm, which can physically filter and intercept large particle impurities and allow the solution containing plasmid DNA to pass through, achieving the purpose of removing large particle impurities and preliminarily purifying the sample.

[0026] After impurity filtration, the bacterial solution flows downward through the filter column 8 into the second column passage 11. In the second column passage 11, plasmid DNA in the bacterial solution is adsorbed onto the adsorption column 10, while the bacterial solution without plasmid DNA flows downward through the adsorption column 10 into the EP tube in the cooling tank 5. This waste liquid can be directly collected in the EP tube and discarded. In the plasmid extraction experiment, the main function of the adsorption column 10 is to specifically bind plasmid DNA, separating it from impurities such as RNA and proteins. Its pore size is usually 2~10μm, which allows plasmid DNA to bind to the silica membrane or similar material inside the column under high salt conditions. After washing to remove impurities, the purified plasmid DNA is obtained by elution under low salt conditions, achieving the purpose of specific binding and purification of plasmid DNA.

[0027] Next, a new EP tube is placed into the cooling bath 5, and two wash solutions are added to the second column passage 11 through the sample addition tube 14. This washes off the plasmid DNA adsorbed on the adsorption column 10 and collects it in the EP tube in the cooling bath 5, completing the plasmid DNA extraction and column passage operation. At the same time, the EP tube is also cooled by the cooling unit 4 on the controller 1, keeping the plasmid DNA in a low temperature environment of 4°C, which helps to increase the stability of the plasmid DNA.

[0028] The entire device can easily perform filtration and adsorption operations on plasmid extraction lysate. A high-pressure air pump 15 can blow nitrogen into the first column passage 9 or the second column passage 11, thereby accelerating the liquid flow through the filter column 8 or adsorption column 10 by pressurizing with high-pressure nitrogen. This ensures no liquid contamination while significantly increasing the liquid flow rate during plasmid extraction. Compared to traditional centrifugation, this method greatly shortens the operation time, avoids plasmid structure damage, and ensures the integrity and function of the final extracted plasmid remain unaffected, improving the success rate and reliability of subsequent experiments. Simultaneously, the final liquid after plasmid extraction is collected in an EP tube within the cooling tank 5, allowing for continuous cooling and effectively increasing plasmid stability, further guaranteeing the quality and activity of the extracted plasmid. Furthermore, the entire device simplifies the cumbersome operation of repeatedly loading and unloading the centrifuge in traditional methods, reducing experimental steps, lowering operational difficulty, and improving experimental efficiency, making it suitable for large-scale plasmid extraction experiments.

[0029] It should be noted that all directional indicators (such as up, down, left, right, front, back, etc.) in this utility model embodiment are only used to explain the relative positional relationship and movement of each component in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicator will also change accordingly.

[0030] In this utility model, unless otherwise explicitly specified and limited, the terms "connection," "fixing," etc., should be interpreted broadly. For example, "fixing" can mean a fixed connection, a detachable connection, or an integral part; it can mean a mechanical connection or an electrical connection; it can mean a direct connection or an indirect connection through an intermediate medium; it can mean the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0031] Furthermore, in this utility model, descriptions involving "first," "second," etc., are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature.

Claims

1. A plasmid extraction column chromatography device, characterized in that: The device includes a controller with a cooling unit. A pressure plate is connected to the controller via a pair of vertically arranged multi-segment telescopic rods. Multiple cooling slots for inserting EP tubes are evenly spaced on the controller between the two multi-segment telescopic rods. A first and second layer plate, directly above the cooling slots, are stacked between the pressure plate and the controller. The first layer plate has multiple first column passages with built-in filter columns, and the second layer plate has multiple second column passages with built-in adsorption columns. The first and second column passages are of the same size. The lower end of the first column passage is a first conical liquid outlet corresponding to the second column passage, and the lower end of the second column passage is a second conical liquid outlet corresponding to the cooling slot. The pressure plate has sample tubes corresponding to the first column passages. The controller contains a high-pressure gas pump for blowing nitrogen gas. The high-pressure gas pump has a blowing pipe extending outside the controller, and the outlet end of the blowing pipe has a branch pipe corresponding to each sample tube.

2. The plasmid extraction column apparatus according to claim 1, characterized in that: The bottom surface of the pressure plate is evenly distributed with a plurality of sealing rings that are correspondingly fastened to the upper end of each of the first column passages, and the sample tubes are respectively sealed and inserted into the pressure plate on the inner side of each of the sealing rings.

3. The plasmid extraction column apparatus according to claim 1 or 2, characterized in that: Electromagnets are provided on the left and right sides of the controller, and adsorption magnets adapted to the electromagnets are provided on the left and right sides of the bottom surface of the pressure plate.

4. The plasmid extraction column apparatus according to claim 3, characterized in that: The front wall of the controller is equipped with a touch screen timer, an air pump switch, and an electromagnetic switch.

5. The plasmid extraction column apparatus according to claim 1, characterized in that: A lifting handle is provided on the top wall of the pressure plate.

6. The plasmid extraction column apparatus according to claim 1, characterized in that: Each of the sample dispensing tubes is equipped with an on / off valve, and the gas outlet branch is connected to the sample dispensing tube on the outlet side of the on / off valve.

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