CN217149150U - Plasmid purification device - Google Patents
Plasmid purification device Download PDFInfo
- Publication number
- CN217149150U CN217149150U CN202221130299.9U CN202221130299U CN217149150U CN 217149150 U CN217149150 U CN 217149150U CN 202221130299 U CN202221130299 U CN 202221130299U CN 217149150 U CN217149150 U CN 217149150U
- Authority
- CN
- China
- Prior art keywords
- eluent
- box
- connecting pipe
- communicated
- plasmid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The utility model provides a plasmid purification device relates to molecular biotechnology field, the utility model provides a pair of plasmid purification device includes: sample case, conveying part, chromatographic column, connecting pipe, nucleic acid detection part, waste liquid case, finished product case and a plurality of eluent storage tanks. The connecting pipe is communicated with the sample box and the input port of the conveying component, and the output port of the conveying component is communicated with the inlet of the chromatographic column. The eluent is stored in each eluent storage box, and the concentrations of the eluents are different; each eluent storage box is communicated with the connecting pipe. The waste liquid box and the finished product box are communicated with the outlet of the chromatographic column through the nucleic acid detection component. Separating impurities in the plasmid sample by arranging a chromatographic column, and collecting the purified plasmid by eluent in an eluent storage box to improve the purity of the plasmid; and moreover, a plurality of eluents with different concentrations are arranged, so that plasmid samples with different purities are obtained, and the experimental requirements are met.
Description
Technical Field
The utility model belongs to the technical field of molecular biotechnology and specifically relates to a plasmid purification device is related to.
Background
Plasmids are circular double-stranded DNA molecules that exist extrachromosomally in bacteria, are capable of independent replication, and are stably inherited. The plasmid is a common carrier of genetic engineering, and can carry exogenous genes into a host for amplification and expression. The purity of the plasmid is directly related to the success of the subsequent experiments. Therefore, it is of great significance to extract high purity plasmids from bacterial cells.
The traditional alkaline lysis method is the most common method for extracting plasmids, and achieves the separation purpose through the denaturation and renaturation difference of chromosome DNA and plasmid DNA. Under conditions of high pH, the hydrogen bonds of DNA break and the double helix structure unravels, while the two complementary strands of the supercoiled covalently closed circular plasmid DNA do not completely separate. When the pH value is adjusted to be neutral by using a Kac solution with low pH value, the deformed plasmid DNA is restored to the original configuration and is stored in the solution, and the chromosome DNA cannot be renatured.
However, the plasmid obtained by the alkaline cleavage method contains more impurities such as protein, RNA, denatured supercoiled plasmid and the like, so that the purity of the plasmid is not high, and the plasmid has certain influence on the success of the experiment.
SUMMERY OF THE UTILITY MODEL
The utility model discloses (one) the problem that will solve is: the plasmid obtained by the alkaline cracking method has low purity, and the success rate of the experiment is influenced.
(II) technical scheme
In order to solve the technical problem, the embodiment of the utility model provides a plasmid purification device, include: the device comprises a sample box, a conveying part, a chromatographic column, a connecting pipe, a nucleic acid detection part, a waste liquid box, a finished product box and a plurality of eluent storage boxes;
the connecting pipe is communicated with the sample box and the input port of the conveying component, and the output port of the conveying component is communicated with the inlet of the chromatographic column;
eluent is stored in each eluent storage box, and the concentration of the eluent is different; each eluent storage box is communicated with the connecting pipe, and a first check valve is arranged between each eluent storage box and the connecting pipe;
the waste liquid box and the finished product box are communicated with an outlet of the chromatographic column through the nucleic acid detection part.
According to an embodiment of the present invention, further, a plurality of first openings are formed on a side wall of the connecting pipe, and the first openings are disposed in one-to-one correspondence with the eluent storage box;
the eluent storage tank is in communication with the first opening, and the first flow-stop valve is located between the first opening and the eluent storage tank.
According to an embodiment of the present invention, further, the packing of the chromatography column is anion exchange resin.
According to an embodiment of the present invention, the apparatus further comprises a balance liquid storage tank, wherein balance liquid is stored in the balance liquid storage tank;
the connecting pipe is provided with a second opening, the balancing liquid storage tank is communicated with the second opening, and a second check valve is arranged between the balancing liquid storage tank and the second opening.
According to an embodiment of the utility model, further, still include and wash miscellaneous liquid storage box;
impurity washing liquid is stored in the impurity washing liquid storage box, a third opening is formed in the side wall of the connecting pipe, and the impurity washing liquid storage box is communicated with the third opening; and a third check valve is arranged between the impurity washing liquid storage tank and the connecting pipe.
According to an embodiment of the present invention, further, the nucleic acid detecting part is an ultraviolet detector.
According to an embodiment of the present invention, further, the conveying member is a peristaltic pump.
According to an embodiment of the present invention, further, the outlet of the sample box is provided with a fourth flow stop valve.
According to an embodiment of the present invention, further, the device further comprises a mounting bracket;
the mounting bracket is provided with a clamping ring, and the chromatographic column is connected with the clamping ring.
According to an embodiment of the present invention, further, a fifth stop valve is disposed between the ultraviolet detector and the waste liquid tank;
and a sixth stop valve is arranged between the ultraviolet detector and the finished product box.
The utility model has the advantages that:
the utility model provides a pair of plasmid purification device, include: sample case, conveying part, chromatographic column, connecting pipe, nucleic acid detection part, waste liquid case, finished product case and a plurality of eluent storage tanks. The connecting pipe is communicated with the sample box and the input port of the conveying component, and the output port of the conveying component is communicated with the inlet of the chromatographic column. The eluent is stored in each eluent storage box, and the concentrations of the eluents are different; each eluent storage box is communicated with the connecting pipe, and a first flow stopping valve is arranged between each eluent storage box and the connecting pipe. The waste liquid box and the finished product box are communicated with the outlet of the chromatographic column through the nucleic acid detection component.
Separating impurities in the plasmid sample by arranging a chromatographic column, and collecting the purified plasmid by eluent in an eluent storage box to improve the purity of the plasmid; and, be provided with a plurality of eluant storage boxes to the corresponding eluant that stores multiple concentration for obtain the plasmid sample of different purities, satisfy the experiment demand.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the technical solutions in the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a plasmid purification apparatus provided by an embodiment of the present invention.
Icon: 11-sample box; 12-a connecting tube; 13-a fourth stop valve;
21-a conveying member;
31-a chromatography column;
41-nucleic acid detecting means;
51-waste liquid tank; 52-a fifth stop valve;
61-eluent storage box; 62-a first check valve;
71-an equilibrating fluid storage tank; 72-a second check valve;
81-impurity washing liquid storage box; 82-third stop valve
91-finished product box; 92-sixth stop valve.
Detailed Description
The technical solution of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
It should be noted that in the description of the present invention, the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplification of description, but 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 thus, should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
It should be noted that, in the description of the present invention, the terms "connected" and "mounted" should be interpreted broadly, for example, they may be fixedly connected, detachably connected, or integrally connected; can be directly connected or connected through an intermediate medium; either mechanically or electrically. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.
In addition, it should be noted that the experimental methods, detection methods, and devices disclosed in the present invention all employ conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields.
The embodiment of the utility model provides a plasmid purification device for improve the purity of plasmid, as shown in figure 1, plasmid purification device includes: sample tank 11, transport unit 21, chromatography column 31, nucleic acid detection unit 41, waste liquid tank 51, product tank 91, and a plurality of eluent storage tanks 61.
Wherein, the sample box 11 is used for storing plasmid samples to be purified. Which communicates with the chromatography column 31 through the connection tube 12. A transport unit 21 for transporting the plasmid to be purified in the sample tank 11 into the chromatography column 31; the conveying part 21 comprises an input port and an output port, one end of the connecting pipe 12 is communicated with the input port of the conveying part 21, and the other end of the connecting pipe is communicated with the sample box 11; the output port of the transport unit 21 communicates with the chromatography column 31 through a pipe. And a waste liquid tank 51 for collecting waste liquid generated in the purification process. And a finished product box 91 for collecting the plasmids after the impurities are removed. A chromatographic column 31 for separating impurities in the sample to be purified. And an eluent for bringing the sample separated from the impurities in the chromatographic column 31 into the finished product tank 91. And a nucleic acid detecting part 41 for monitoring the loading process and the elution process.
The embodiment of the utility model provides a plasmid purification device, including a plurality of eluant storage boxes 61 for the sample in elution chromatographic column 31, with the plasmid sample that obtains different purities.
As shown in fig. 1, each eluent storage tank 61 stores an eluent, and the concentration of the eluent in each eluent storage tank 61 is uniform. Optionally, the concentration of the eluent can be set up in a proportion increasing/decreasing manner, and the required concentration of the eluent can also be set up in combination with the practical situation of the experiment. All eluent storage tanks 61 are communicated with the connecting pipe 12. The plasmid purification device in this embodiment further includes a first check valve 62, the first check valve 62 is disposed in one-to-one correspondence with the eluent storage tank 61, and the first check valve 62 is disposed between the eluent storage tank 61 and the connection pipe 12, and is used for controlling the opening or closing of the corresponding eluent storage tank 61.
The outlet of the chromatography liquid is connected to the nucleic acid detecting unit 41, and the waste liquid tank 51 and the finished product tank 91 are connected to the outlet of the chromatography column 31 via the nucleic acid detecting unit 41.
In this embodiment, the purity of plasmid is improved by arranging the chromatographic column 31 to separate impurities in the plasmid sample and collecting purified plasmid through the eluent in the eluent storage box 61; and, be provided with a plurality of eluant storage tanks 61 to the corresponding eluant that stores multiple concentration for obtain the plasmid sample of different purities, satisfy the experiment demand.
In the present embodiment, as shown in fig. 1, the eluent storage tank 61 and the connection pipe 12 are connected to each other by opening the connection pipe 12.
Specifically, the connecting pipe 12 is provided with a plurality of first openings, and the first openings are arranged in one-to-one correspondence with the eluent storage boxes 61. Eluent storage tank 61 communicates with the first opening, and first check valve 62 is disposed between the first opening and eluent storage tank 61.
Optionally, in this embodiment, the eluent storage tank 61 and the first opening may be connected through the first check valve 62, or may be connected through a pipeline. When the first opening is connected to the eluent storage box 61 through a pipeline, the first check valve 62 is disposed on the pipeline for controlling the on/off of the pipeline.
Alternatively, in this embodiment, the eluent storage tank 61 can be connected to the connection pipe 12 through a multi-way joint.
Alternatively, in this embodiment, the number of first openings may be greater than the number of eluent storage tanks 61. When the number of the first openings is larger than that of the eluent storage box 61, a part of the first openings are connected with the eluent storage box 61, and the rest part of the first openings are sealed by the plugs.
In actual use, the packing of the chromatographic column 31 is an anion exchange resin.
In an alternative embodiment of this embodiment, the packing of the column 31 may also be DEAE anion exchange packing.
In this embodiment, as shown in fig. 1, an equilibrium liquid storage tank 71 for storing an equilibrium liquid is further included.
The equilibrium liquid storage tank 71 stores equilibrium liquid. A second opening is provided in the connecting pipe 12, and the equalizing liquid storage tank 71 communicates with the second opening. Alternatively, the equalizing fluid storage tank 71 may be in communication with the second opening via a line, or may be directly connected via a second check valve 72. When the equalizing liquid storage tank 71 is communicated with the second opening through a pipeline, a second check valve 72 is provided on the pipeline for controlling the on-off of the pipeline.
In this embodiment, the equilibration fluid is used to activate the anion exchange resin packing within the chromatography column 31.
In use, an operator starts the peristaltic pump, closes the first check valve 62, the third check valve 82 and the fourth check valve 13, opens the second check valve 72, pumps the balance liquid in the balance liquid storage tank 71 into the chromatographic column 31, and activates the anionic resin; then the second stop valve 72 is closed and the fourth stop valve 13 is opened, all other stop valves being closed, and the sample in the sample tank 11 is pumped into the chromatography column 31. The fourth stop valve 13 is then closed and the third stop valve 82 and corresponding first stop valve 62 are opened, completing the elution.
According to the plasmid purification apparatus provided by this embodiment, as shown in FIG. 1, a washing impurity storage tank 81 for storing washing impurity liquid is further included.
The impurity washing liquid storage tank 81 stores impurity washing liquid. A third opening for communicating with the impurity washing liquid storage tank 81 is opened on the side wall of the connection pipe 12, and the impurity washing liquid storage tank 81 communicates with the third opening. Optionally, the impurity washing liquid storage tanks 81 can be communicated with each other through a pipeline, or can be directly connected through the third check valve 82. When the impurity washing liquid storage tank 81 is communicated with the third opening through a pipeline, the third check valve 82 is arranged on the pipeline and used for controlling the on-off of the pipeline.
In an alternative embodiment of this embodiment, the nucleic acid detecting part 41 is an ultraviolet detector.
In this embodiment, the ultraviolet detector has a display device, and optionally, may be an external computer. The whole elution process was monitored by an ultraviolet detector.
Specifically, the ultraviolet monitor is used for monitoring the purity of the plasmid.
In the present embodiment, as shown in fig. 1, the waste liquid tank 51 and the finished product tank 91 communicate with a pipe from which the ultraviolet detector extends.
Optionally, a three-way joint may be provided at the ultraviolet detector to communicate the waste liquid tank 51 and the finished product tank 91.
In practical use, the ultraviolet detector further comprises a fifth check valve 52 and a sixth check valve 92, wherein the fifth check valve 52 is used for controlling the connection and disconnection between the waste liquid tank 51 and the ultraviolet detector; the sixth check valve 92 is used for controlling the connection and disconnection between the finished product box 91 and the ultraviolet detector.
In use, an operator judges the purity of the plasmid according to the data detected by the ultraviolet detector, and collects the plasmid by controlling the on-off of the fifth check valve 52 and the sixth check valve 92.
Specifically, the fifth check valve 52 is disposed between the waste liquid tank 51 and the three-way joint; the second check valve 72 is provided between the finished product tank 91 and the three-way joint.
Optionally, in this embodiment, the first stop valve 62, the second stop valve 72, the third stop valve 82, the fourth stop valve 13, the fifth stop valve 52 and the sixth stop valve 92 may also be configured to cooperate with a flow-limiting clamp by arranging a hose to replace the functions of the above-mentioned stop valves.
Optionally, in this embodiment, a mounting bracket is further included for mounting the chromatography column 31.
In this embodiment, a snap ring is provided on the mounting bracket, and the chromatography column 31 is connected to the snap ring. Preferably, the chromatographic column 31 is detachably connected with the clamping ring.
Optionally, in this embodiment, the mounting bracket may be an iron stand.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention.
Claims (10)
1. A plasmid purification apparatus, comprising: a sample box (11), a conveying component (21), a chromatographic column (31), a connecting pipe (12), a nucleic acid detection component (41), a waste liquid box (51), a finished product box (91) and a plurality of eluent storage boxes (61);
the connecting pipe (12) is communicated with the sample box (11) and the input port of the conveying component (21), and the output port of the conveying component (21) is communicated with the inlet of the chromatographic column (31);
eluent is stored in each eluent storage box (61) and the concentration of the eluent is different; each eluent storage box (61) is communicated with the connecting pipe (12), and a first check valve (62) is arranged between each eluent storage box (61) and the connecting pipe (12);
the waste liquid tank (51) and the finished product tank (91) are communicated with an outlet of the chromatographic column (31) through the nucleic acid detecting part (41).
2. The plasmid purification device according to claim 1, wherein the side wall of the connecting pipe (12) is provided with a plurality of first openings, and the first openings are arranged in one-to-one correspondence with the eluent storage tanks (61);
the eluent storage tank (61) is in communication with the first opening, and the first flow stop valve (62) is located between the first opening and the eluent storage tank (61).
3. The plasmid purification apparatus according to claim 1, wherein the packing of the chromatography column (31) is an anion exchange resin.
4. The plasmid purification apparatus according to claim 3, further comprising an equilibration fluid storage tank (71), wherein an equilibration fluid is stored in the equilibration fluid storage tank (71);
the connecting pipe (12) is provided with a second opening, the balancing liquid storage tank (71) is communicated with the second opening, and a second check valve (72) is arranged between the balancing liquid storage tank (71) and the second opening.
5. The plasmid purification apparatus according to claim 4, further comprising a wash liquor storage tank (81);
impurity washing liquid is stored in the impurity washing liquid storage box (81), a third opening is formed in the side wall of the connecting pipe (12), and the impurity washing liquid storage box (81) is communicated with the third opening; and a third check valve (82) is arranged between the impurity washing liquid storage tank (81) and the connecting pipe (12).
6. The plasmid purification apparatus according to claim 1, wherein the nucleic acid detection part (41) is an ultraviolet detector.
7. The plasmid purification apparatus according to claim 1, wherein the conveying means (21) is a peristaltic pump.
8. Plasmid purification device according to claim 1, characterized in that the outlet of the sample box (11) is provided with a fourth stop valve (13).
9. The plasmid purification apparatus of claim 1, further comprising a mounting bracket;
the mounting bracket is provided with a clamping ring, and the chromatographic column (31) is connected with the clamping ring.
10. The plasmid purification device according to claim 6, wherein a fifth check valve (52) is arranged between the ultraviolet detector and the waste liquid tank (51);
and a sixth stop valve (92) is arranged between the ultraviolet detector and the finished product box (91).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202221130299.9U CN217149150U (en) | 2022-05-05 | 2022-05-05 | Plasmid purification device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202221130299.9U CN217149150U (en) | 2022-05-05 | 2022-05-05 | Plasmid purification device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN217149150U true CN217149150U (en) | 2022-08-09 |
Family
ID=82665501
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202221130299.9U Active CN217149150U (en) | 2022-05-05 | 2022-05-05 | Plasmid purification device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN217149150U (en) |
-
2022
- 2022-05-05 CN CN202221130299.9U patent/CN217149150U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5330914A (en) | Apparatus and method for automated purification of extra-chromo-somal DNA from a cell suspension in a container | |
| US9506847B2 (en) | Method and system for selective isolation of target biological molecules in a general purpose system | |
| EP2675540B1 (en) | System and process for biopolymer chromatography | |
| US5346999A (en) | Method of nucleic acid extraction | |
| JP2017508143A (en) | Automated multi-stage purification system | |
| CN107485891B (en) | Improved chromatographic device and method for continuous flow chromatography | |
| CN217149150U (en) | Plasmid purification device | |
| US10843104B2 (en) | System and process for biopolymer chromatography | |
| CA1335963C (en) | Automated nucleic acid extractor | |
| Smith et al. | Hydrophobic interaction ligand selection and scale-up of an expanded bed separation of an intracellular enzyme from Saccharomyces cerevisiae | |
| CN205653449U (en) | Simple and easy device with negative pressure method purification nucleic acid | |
| CN218058999U (en) | Protein fermentation liquor pretreatment device for recombinant escherichia coli expression system | |
| JPH04505848A (en) | Apparatus and method for automatic purification of extrachromosomal DNA from cell suspension | |
| CN219308758U (en) | Microfluidic protein purification core, detection system and double-purification unit chip | |
| CN215799619U (en) | Cell culture equipment | |
| CN201280551Y (en) | System for purifying bacteria magnetosome | |
| CN105675864A (en) | Automatic bacteria sorting and marking device based on immune method | |
| CN115747042A (en) | Full-automatic integrated nucleic acid extraction equipment for large-volume samples | |
| CN113265396B (en) | Process for continuous production of large plasmid DNA | |
| CN215799464U (en) | Adenovirus purification device | |
| CN216170113U (en) | Instrument for realizing automatic affinity chromatography | |
| CN216630346U (en) | Tangential flow ultrafiltration system | |
| CN219050408U (en) | Protein purification system | |
| JP2003530868A (en) | MIPC column cleaning system and method | |
| CN210834791U (en) | Online solvent recovery unit based on detector |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |