CN112210476A - Be applied to biological liquid workstation - Google Patents
Be applied to biological liquid workstation Download PDFInfo
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- CN112210476A CN112210476A CN202011261784.5A CN202011261784A CN112210476A CN 112210476 A CN112210476 A CN 112210476A CN 202011261784 A CN202011261784 A CN 202011261784A CN 112210476 A CN112210476 A CN 112210476A
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- 239000007788 liquid Substances 0.000 title claims abstract description 68
- 239000011324 bead Substances 0.000 claims abstract description 69
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 65
- 238000010438 heat treatment Methods 0.000 claims abstract description 30
- 230000007246 mechanism Effects 0.000 claims abstract description 28
- 239000002699 waste material Substances 0.000 claims description 30
- 238000012546 transfer Methods 0.000 claims description 25
- 230000010355 oscillation Effects 0.000 claims description 20
- 238000002156 mixing Methods 0.000 claims description 18
- 238000011084 recovery Methods 0.000 claims description 8
- 230000005540 biological transmission Effects 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 238000003780 insertion Methods 0.000 claims 2
- 230000037431 insertion Effects 0.000 claims 2
- 239000013060 biological fluid Substances 0.000 claims 1
- 150000007523 nucleic acids Chemical class 0.000 abstract description 11
- 102000039446 nucleic acids Human genes 0.000 abstract description 11
- 108020004707 nucleic acids Proteins 0.000 abstract description 11
- 238000000605 extraction Methods 0.000 abstract description 10
- 238000000926 separation method Methods 0.000 description 22
- 239000000523 sample Substances 0.000 description 18
- 238000000034 method Methods 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 238000010276 construction Methods 0.000 description 3
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- 239000013076 target substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 239000005441 aurora Substances 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
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- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
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- 238000007599 discharging Methods 0.000 description 1
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- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000005213 imbibition Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
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- 239000000696 magnetic material Substances 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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Abstract
The invention discloses a liquid workstation applied to biology, which comprises a working platform, a working disk surface, a pipetting arm and a pipetting frame, wherein the pipetting arm is arranged right above the working disk surface through an x-axis driving mechanism, a y-axis driving mechanism and a Z-axis driving mechanism, the x-axis of the pipetting arm is parallel to an equipment box, the area of the working disk surface comprises nine working disk positions which are arranged in a Sudoku mode, and a first gun head adapter, a second gun head adapter, a sample collecting carrier, a third gun head adapter, a reagent tank carrier, a temperature control adapter, a magnetic bead mixer, a magnetic bead separating disk and a heating and vibrating disk are sequentially arranged on the working disk positions from inside to outside and from left to right; the invention has the advantages of compact overall structure layout, adoption of an open kit, suitability for kits of various manufacturers, high automation of the extraction process, avoidance of pollution, improvement of extraction efficiency, and capability of purifying and extracting macromolecular products such as high-quality and high-purity nucleic acid.
Description
Technical Field
The invention relates to the field of biology, in particular to a liquid workstation applied to biology.
Background
With the rapid development of life science and technology, the research work of biochemical laboratories is faced with more complicated research objects and increasing sample quantities, the traditional manual operation mode cannot meet the requirements of high efficiency, accuracy and safety in the high-throughput sample processing process, and the automatic operation of part or the whole process in the experimental operation flow becomes a necessary trend.
Nucleic acid is the basic object of molecular biology research, and the extraction of nucleic acid is the most important and basic link in molecular diagnosis. There are many methods for extracting DNA or RNA from a biological sample, such as a conventional organic solvent extraction method, and a silica gel column adsorption method, a charge method, and a magnetic bead separation method, which are suitable for a 96-well plate format for high-throughput automation. Among them, the magnetic bead separation method is currently the most widely used method with the most mature automation conditions. The magnetic bead is a novel functionalized solid-phase carrier, has active groups on the surface, can be coupled with various substances, has the characteristics of liquid fluidity and solid magnetic materials, and can directionally move and concentrate under the action of an external magnetic field. When the external field magnetic field is removed, the target substance can be uniformly dispersed in the liquid by slight oscillation or suction, so that the separation of the solid phase and the liquid phase becomes very quick and convenient, and the target substance with high purity can be obtained by simple elution.
In the prior art, the automatic liquid treatment workstation and the liquid transfer device therein mostly adopt a sliding friction pair mechanism to complete the movement of the liquid transfer suction head in the vertical direction, the mechanism has large abrasion, large noise and low movement stability, the processing precision requirement of a sliding chute of a movable base is higher, and the guarantee is difficult. In addition, in the pipetting device in the prior art, the movable base of the pipetting mechanism is positioned in the vertical direction of the rotating shaft and is mostly restrained by the combination of the spline and the roller, the debugging is complicated in the assembling process, and the workload is large.
In biological experiments, for example, the establishment of a reaction system for nucleic acid extraction and PCR generally requires multiple pipetting, the experiment operation is complicated, the extraction efficiency of a sample is limited, and nucleic acid extracted by the existing magnetic bead method nucleic acid extraction method is mixed with more impurities and needs to be repeatedly purified again to meet the requirements. And manual operation is adopted, so that pollution among samples is easily caused.
Therefore, in view of the above problems, there is a need for a liquid biostation apparatus that can improve the efficiency of nucleic acid extraction and reduce the problem of nucleic acid contamination.
Disclosure of Invention
The present invention is directed to a liquid workstation for biological applications, which solves the above-mentioned problems of the prior art.
In order to achieve the purpose, the invention provides the following technical scheme:
a liquid workstation applied to biology comprises a working platform, a working disk surface, a pipetting arm and a pipetting frame, wherein the working disk surface is arranged on the working platform, an equipment box is vertically arranged on the rear top surface of the working platform, the pipetting arm is arranged at the top of the equipment box through an x-axis driving mechanism, a y-axis driving mechanism and a Z-axis driving mechanism, the x-axis of the pipetting arm is parallel to the equipment box, the working disk surface area comprises nine working disk positions which are arranged in a nine-palace-grid mode, and a first gun head adapter, a second gun head adapter, a sample collecting carrier, a third gun head adapter, a reagent tank carrier, a temperature control adapter, a magnetic bead mixer, a magnetic bead separating disk and a heating vibration disk are sequentially arranged in the working disk positions from inside to outside and from left to;
the multi-specification gun heads are placed on the first gun head adapter, the second gun head adapter and the third gun head adapter, a sucker mechanism for sucking the gun heads on the first gun head adapter, the second gun head adapter and the third gun head adapter is arranged on the liquid transfer arm, a standard pore plate for placing an experimental sample to be tested is arranged on the sample collection carrier, multi-specification reagent bottles are placed on the reagent tank carrier, multi-specification PCR tubes are placed on the top of the temperature control adapter, a temperature control device for heating and refrigerating the PCR tubes is arranged at the bottom of the temperature control adapter, and a waste area is arranged on the left side of the magnetic bead mixer;
the magnetic bead separating disc comprises a magnetic rod, the heating oscillation disc comprises a bottom vortex oscillation mechanism, a moving disc frame is arranged on the magnetic bead separating disc or the heating oscillation disc, a centrifugal tube disc is arranged on the moving disc frame, the moving disc frame is arranged on a working disc position on a working disc surface, and a hook lifting mechanism for lifting the moving disc frame is arranged on the moving arm.
Further scheme: the centrifuge tube disk is provided with a centrifuge tube placing area for placing the centrifuge tube body and a centrifuge tube fixing area for fixing the centrifuge tube cover body.
Further scheme: move a set frame and include solution dish carriage and move a set hook frame, solution dish carriage is put on the shelf and is being put on the centrifuge tube dish, and solution dish carriage support frame is established around the work position opening of work quotation, and work position opening edge is equipped with and is used for erectting the frame groove that is equipped with the support solution dish carriage, move a set hook frame and vertically install on the lateral wall of solution dish carriage.
Further scheme: move the lift seat that vertical direction removed is installed to the Z axle bottom of liquid arm, the hook is got the mechanism and is included the hook piece of installing on lift seat and install the couple hole piece on moving a set hook frame top lateral wall, vertically on the hook piece top surface be fixed with a plurality of hook nails, a plurality of nail holes that are used for inserting the hook nail are seted up to hook piece bottom surface.
Further scheme: the waste object area comprises a waste liquid groove, a waste removing suction head device and a gun head recovery box, the gun head recovery box is arranged at the bottom of the working disk surface and is located below the waste removing suction head device, the suction head device comprises a gun head plugging rod arranged at the bottom of a Z shaft of a liquid transfer arm, the waste removing suction head device comprises a suction head rack, and an opening hole used for clamping the gun head plugging rod is formed in a top plate of the suction head rack.
Further scheme: and a through air channel is arranged in the gun head inserting rod, one end of the air channel is opened at the bottom end of the gun head inserting rod and is used for generating negative pressure liquid suction to the gun head, and the other end of the air channel is communicated with a gas pumping and discharging end of a liquid pump arranged in the equipment box through an air pipe.
Further scheme: the whole vertical fixing of bar magnet is on the bar magnet base, and bar magnet base fixed mounting is in work platform, and the installation of bar magnet is located and is relative with the tube hole position of centrifuging tube on the centrifuging tube dish.
Further scheme: the utility model discloses a centrifugal tube box, including working platform, worm-screw vibration mechanism motor base, vibrate pipe dish and shock motor, motor base fixed mounting is in work platform, and the motor that vibrates is installed at motor base bottom surface center, and the motor shaft that vibrates the motor has a vibration pipe dish through eccentric cam piece horizontal mounting, vibrates the pipe dish top surface and has seted up a plurality of body grooves that are used for placing the centrifuging tube, and the position that the centrifuging tube box placed through moving the dish frame and corresponding the centrifuging tube is laid on the body groove that vibrates the pipe dish, the bottom of vibrating the pipe dish is provided with the heating film.
Further scheme: the magnetic bead mixer comprises a magnetic bead container, a container bearing device, an eccentric vibrator, a buffer device and a mixing motor, wherein the magnetic bead container is sleeved in the container bearing device, the center of the bottom of the container bearing device is connected with the motor shaft of the mixing motor through the eccentric vibrator in a transmission mode, and the buffer device used for limiting the left-right swinging amplitude of the container bearing device is arranged on two sides of the bottom surface of the container bearing device.
Compared with the prior art, the invention has the beneficial effects that: the invention has the advantages that the overall structure layout is very compact, the components can not interfere with each other or collide with each other when executing respective tasks, an open kit is adopted, the kit is suitable for kits of various manufacturers, the extraction process is highly automated, pollution is avoided, the extraction efficiency is improved, and macromolecular products such as high-quality and high-purity nucleic acid can be purified and extracted.
Drawings
FIG. 1 is a schematic view of a liquid workstation applied to biology.
Fig. 2 is a schematic view of mounting position structures of a first gun head adapter, a second gun head adapter, a sample collection carrier, a third gun head adapter, a reagent tank carrier, a temperature control adapter, a magnetic bead mixer, a magnetic bead separation disc and a heating oscillation disc in a liquid workstation applied to biology.
FIG. 3 is a schematic diagram of a pipetting arm in a fluid station for biological applications.
FIG. 4 is a schematic diagram of the structure of the work plate surface in the liquid work station applied to biology.
FIG. 5 is a schematic view showing the structure of a reagent tank carrier used in a liquid work station for biology.
Fig. 6 is an exploded view of a magnetic bead separation disk applied to a liquid work station in biology.
FIG. 7 is an exploded view of a heated vibrating plate used in a liquid workstation for biology.
FIG. 8 is a schematic diagram of a magnetic bead mixer used in a liquid workstation for biology.
In the figure: the pipette tip device comprises a pipette arm 1, a tray moving rack 2, a tray moving hook rack 20, a hook hole block 21, a support frame 22, a work disc surface 3, a waste liquid tank 301, a waste tip removing device 302, a tip recovery box 303, a first tip adapter 31, a second tip adapter 32, a sample collection carrier 33, a third tip adapter 34, a reagent tank carrier 35, a temperature control adapter 36, a magnetic bead separation disc 37, a magnetic rod 373, a magnetic rod base 374, a heating oscillation disc 38, a motor base 382, an oscillation motor 383, an eccentric cam block 384, a centrifuge tube disc 39, a centrifuge tube placing area 390, a centrifuge tube fixing area 391, a magnetic bead mixer 4, a magnetic bead container 40, a container carrying device 41, an eccentric vibrator 42, a buffer device 43, a mixing motor 44, a work platform 6 and an equipment box 60.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Referring to fig. 1 to 8, in the embodiment of the present invention, a liquid workstation applied to biology includes a working platform 6, a working disk surface 3, a pipetting arm 1, and a pipetting frame 2, where the working disk surface 3 is disposed on the working platform 6, an equipment box 60 is vertically installed on a rear top surface of the working platform 6, the pipetting arm 1 is installed on a top of the equipment box 60 through an x-axis, a y-axis, and a Z-axis driving mechanism, an x-axis of the pipetting arm 1 is parallel to the equipment box 60, the area of the working disk surface 3 includes nine working disk positions arranged in a grid pattern, and a first gun head adapter 31, a second gun head adapter 32, a sample collection carrier 33, a third gun head adapter 34, a reagent tank carrier 35, a temperature control adapter 36, a magnetic bead mixer 4, a magnetic bead separation disk 37, and a heating oscillation disk 38 are sequentially placed from inside to outside and from left to right;
multiple-specification tips are placed on the first, second and third tip adapters 31, 32 and 34, a pipette tip mechanism for sucking tips on the first, second and third tip adapters 31, 32 and 34 is arranged on the pipette arm 1, a standard pore plate for placing an experimental sample to be tested is arranged on the sample collection carrier 33, multiple-specification reagent bottles are placed on the reagent tank carrier 35, multiple-specification PCR tubes are placed on the top of the temperature control adapter 36, a temperature control device for heating and refrigerating the PCR tubes is arranged at the bottom of the temperature control adapter 36, and a waste area is arranged on the left side of the magnetic bead mixer 4;
the magnetic bead separating disc 37 comprises a magnetic rod 373, the heating oscillation disc 38 comprises a bottom vortex oscillation mechanism, the magnetic bead separating disc 37 or the heating oscillation disc 38 is provided with a disc moving frame 2 in an erected mode, the disc moving frame 2 is provided with a centrifugal tube disc 39 in an erected mode, the disc moving frame 2 is arranged on a working disc position on a working disc surface 3 in an erected mode, and the transfer arm 1 is provided with a hook taking mechanism used for lifting the whole disc moving frame 2.
The centrifuge tube tray 39 is provided with a centrifuge tube placing area 390 for placing the centrifuge tube body and a centrifuge tube fixing area 391 for fixing the centrifuge tube cover body.
The moving disc frame 2 comprises a solution disc supporting frame 22 and a moving disc hook frame 20, a centrifugal tube disc 39 is erected on the solution disc supporting frame 22, the solution disc supporting frame 22 supports the frame and is arranged around a working disc position opening of the working disc surface 3, a frame groove used for supporting the solution disc supporting frame 22 is formed in the edge of the working disc position opening, and the moving disc hook frame 20 is vertically arranged on the side wall of the solution disc supporting frame 22.
The lifting seat 10 that vertical direction removed is installed to the Z axle bottom of pipetting arm 1, the mechanism is got including installing gib head piece 11 on lifting seat 10 and installing the couple hole piece 21 on moving a set of hook frame 20 top lateral wall to the hook, vertically on the gib head piece 11 top surface be fixed with a plurality of hook nails, a plurality of nail holes that are used for inserting the hook nail are seted up to hook piece 21 bottom surface, and the pipetting arm 1 that sets up can directly upwards mention moving a set of 2 through mutually supporting of gib head piece 11 and couple hole piece 21, then supports centrifuge tube dish 39 on moving a set of 2 alright remove on the working dish position.
The waste object area comprises a waste liquid groove 301, a waste suction head removing device 302 and a gun head recovery box 303, the gun head recovery box 303 is arranged at the bottom of the working disk surface 3 and is located below the waste suction head removing device 302, the suction head device comprises a gun head plugging rod 12 arranged at the bottom of a Z shaft of a pipetting arm 1, the waste suction head removing device 302 comprises a suction head frame, an opening hole used for clamping the gun head plugging rod 12 is formed in a top plate of the suction head frame, when the gun head plugging rod 12 drives a gun head of a socket to move to one side of the suction head frame, the rod wall of the gun head plugging rod 12 at the top of the gun head transversely slides into the opening hole by the pipetting arm 1, the top end of the gun head is located on the bottom surface of the opening hole, the pipetting arm 1 is lifted upwards again, the gun head is tripped from the gun head plugging rod 12 and.
The automatic liquid suction gun is characterized in that a through air path channel is arranged in the gun head plugging rod 12, one end of the air path channel is used for generating negative pressure liquid suction to the gun head at an opening at the bottom end of the gun head plugging rod 12, the other end of the air path channel is communicated with a gas suction and discharge end of a liquid pump which is arranged in the equipment box 60 through an air pipe, and after the gun head is plugged by the arranged gun head plugging rod 12, the liquid pump can perform washing liquid or liquid drainage operation on the gun head through the gun head plugging rod 12.
The magnetic rod 373 is integrally and vertically fixed on the magnetic rod base 374, the magnetic rod base 374 is fixedly installed in the working platform 6, the magnetic rod 373 is installed in a position opposite to the tube hole of the centrifugal tube on the centrifugal tube plate 39, and the magnetic rod 373 is used for performing magnetic bead separation operation on the centrifugal tube in the tube hole of the centrifugal tube plate 39.
The utility model discloses a quick vibration of spiral vibration mechanism motor base 382, vibrate pipe dish 381 and vibrate motor 383, motor base 382 fixed mounting is in work platform 6, and motor base 382 installs in the bottom surface center and vibrates motor 383, and the motor shaft that vibrates motor 383 has a vibration pipe dish 381 through eccentric cam piece 384 horizontal installation, vibrates the pipe dish 381 top surface and has seted up a plurality of body grooves that are used for placing the centrifuging tube, and the position that centrifugal tube box 39 placed through moving the dish frame 2 and corresponding the centrifuging tube is laid on the body groove that vibrates pipe dish 381, the bottom that vibrates pipe dish 381 is provided with the heating film, and the vibration pipe dish 381 that sets up can be under the drive of vibrating motor 383 and eccentric cam piece 384, at the eccentric swing of horizontal direction, realizes the action of quick vibration, can carry out quick vibration operation to the centrifugal tube dish 39 of placing above.
The magnetic bead mixer 4 comprises a magnetic bead container 40, a container bearing device 41, an eccentric vibrator 42, a buffer device 43 and a mixing motor 44, wherein the magnetic bead container 40 is sleeved in the container bearing device 41, the center of the bottom of the container bearing device 41 is connected with the motor shaft of the mixing motor 44 through the eccentric vibrator 42 in a transmission manner, the buffer devices 43 used for limiting the left-right swinging amplitude of the container bearing device 41 are arranged on two sides of the bottom surface of the container bearing device 41, after the magnetic bead container 40 is arranged in the container bearing device 41, the mixing motor 44 can drive the container bearing device 41 to horizontally swing through the eccentric vibrator 42, and the magnetic bead container 40 can perform rapid mixing operation on magnetic beads.
Example 2
This example differs from example 1 in that: the first lance tip adapter 31, the second lance tip adapter 32 and the third lance tip adapter 34 can be used for placing lance tips with the specifications of 1000, 200, 100 and 50 μ L according to different experiments. Preferably, a gun head with the specification of 1000 mu L is placed on the first gun head adapter 31, a gun head with the specification of 200 mu L is placed on the second gun head adapter 32, and a gun head with the specification of 1000 mu L is placed on the third gun head adapter 34;
wherein the sample collection carrier 33 in the work surface 3 is used for placing the experimental to-be-tested object, and the sample collection carrier 33 is not limited to a standard 24-well plate, a standard 48-well plate, a standard 96-well plate and the like. Preferably, we place 24 EP centrifuge tubes on the sample collection carrier 33.
In the reagent tank carrier 35 in the working disc surface 3 of the present embodiment, considering that the use habit of the user is that the reagent bottles are put into the instrument in a whole bottle, preferably, 82 mL tubes are adapted; 4 50mL tubes.
The temperature control adapter 36 in the working disc surface 3 of this embodiment can be used for temperature control reaction, product storage, etc., the temperature can be controlled to be 4-99 ℃, and preferably, the temperature control adapter can be used for storing 8-joint PCR tubes of a drum cover and is used for configuration of PCR amplification reaction liquid, etc.
The magnetic bead blender 4 in this embodiment working disc face 3, 2 mL's magnetic bead that can be even is applied to the magnetic bead method and draws the nucleic acid field, can make magnetic bead and solution mixing, and it is effectual to mix, and the mixing efficiency is high, makes test sample and magnetic bead even combination, improves the concentration and the purity of extracting.
Magnetic bead separation dish 37 in service plate face 3, can effectively separate magnetic bead and solution, magnetic bead separation dish 37 contains bar magnet 373, and centrifuge tube dish 39 has optimized, has increased centrifuging tube fixed area 391, makes the magnetic bead adsorb in same direction, siphons away the magnetic bead in the prevention and cure imbibition flow, reduces the loss of sample. Preferably, we place a 2mL centrifuge tube in the tube placement area 390 and secure the centrifuge tube lid to the tube securing area 391.
The liquid pump installed in the equipment box 60 can be 25 μ L or 1000 μ L, and the robot arm 1 can move freely in the directions of the X-axis (the axis from left to right of the table), the Y-axis (the axis from front to back of the table), and the Z-axis (the vertical axis above the table). Wherein the moving plate rack 2 of the workstation of the invention can move the components of the workstation (such as a deep hole plate) on X, Y, Z axes. A waste area is provided for placing waste liquid generated in the experiment, such as solid waste and/or liquid waste, and the waste area preferably comprises a waste liquid groove 301, a waste suction head removing device 302 and a gun head recovery box 303.
In the present example, the following test reagents were used as Aurora reagents, and nucleic acid extraction by a magnetic bead purification method and construction of a PCR reaction system were performed.
The specific experimental construction method is as follows:
firstly, preparing reagents and consumables (a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a reagent F, a reagent G, a reagent H, a reagent I, a reagent J, a reagent K, a reagent L, a magnetic bead tube and 24 DNA samples to be processed) in advance; and manually placing the reagent in the corresponding location. Reagent tank carrier 35 positions A, B, C, D, E, F, G, H, I, J, K and L respectively hold reagent A, reagent B, reagent C, reagent D, reagent E, reagent F, reagent G, reagent H, reagent I, reagent J, reagent K, and reagent L. 48 0.2mL drum-lid PCR tubes were placed in temperature-controlled adapter 36 in the worktop panel 3. The magnetic bead mixer 4 is used for placing a magnetic bead tube. The sample collection carriage 33 places 24 DNA samples to be treated. The heated shaker plate 38 places 24 EP centrifuge tube cassettes 39 containing 24 2mL uncapped centrifuge tubes. The first lance tip adapter 31 accommodates a 1000 μ L-sized lance tip, the second lance tip adapter 32 accommodates a 200 μ L-sized lance tip, and the third lance tip adapter 34 accommodates a 1000 μ L-sized lance tip.
The implementation steps of the specific experimental construction are as follows:
1. taking 1000 mu L of gun heads from the first gun head adapter 31 by using a liquid transfer arm 1; aspirating 90. mu.L of reagent A from the A position of the reagent tank carriage 35 and placing it in the front 24 wells of the temperature-controlled adapter 36 drum-lid PCR tube D;
2. taking 200 mu L of gun heads from the second gun head adapter 32 by using a liquid transfer arm 1; sucking 10. mu.L of reagent C from the position C of the reagent tank carriage 35 and placing the reagent C in the front 24 wells of the PCR tube covered by the temperature-controlled adapter 36;
3. taking 200 mu L of gun heads from the second gun head adapter 32 by using a liquid transfer arm 1; sucking 20 μ L of samples from the sample collection carrier 33, putting a total of 24 samples into the temperature control adapter 36, covering the PCR tube with a drum, and blowing, beating and uniformly mixing;
4. taking 200 mu L of gun heads from the second gun head adapter 32 by using a liquid transfer arm 1; sucking 50 μ L of reagent D from the position D of the reagent tank carrier 35, placing the reagent D in the front 24 holes of the PCR tube covered by the temperature control adapter 36, and sealing;
5. the temperature control adapter 36 heats the drum cover 8 connecting pipe to react for 10min at 95 ℃; then cooling to 64 ℃ to react for 60 min;
6. taking 200 mu L of pipette tip by using a pipette arm No. 1 second pipette tip adapter 32, and transferring 120 mu L of reaction system from a temperature control adapter 36 tube to a 2mL centrifuge tube of a centrifuge tube box 39;
7. taking 1000 mu L of pipette tip from the I position of the reagent tank carrier 35 by using a pipette arm 1 at a first pipette tip adapter 31, and placing 700 mu L of reagent I into a 2mL centrifuge tube of a centrifuge tube box 39;
8. using a pipetting arm 1 to pipette 40 microliter of magnetic beads from a magnetic bead mixer 37 on a pipette tip adapter 32 of No. two; placing the mixture into a 2mL centrifuge tube of a centrifuge tube box 39, heating an oscillation disc 38, and uniformly mixing the mixture at room temperature for 10 min.
9. Moving a 2mL centrifugal tube box 39 from the heating oscillation disc 38 to the magnetic bead separation disc 37 by using a disc moving frame 2, and carrying out magnetism suction for 2 min;
10. pipette tip of 1000. mu.L is pipetted into the centrifuge tube cassette 39 by the pipette tip adapter 31 using the pipette arm 1, and the waste liquid is aspirated.
11. Taking 1000 mu L of pipette tip from the first pipette tip adapter 31 by using the pipette arm 1, sucking 600 mu L of reagent K from the K position of the reagent tank carrier 35, and adding the reagent K into a centrifuge tube of the centrifuge tube box 39;
12. the centrifuge tube cassette 39 is moved from the magnetic bead separation disk 37 to the heating oscillation disk 38 by the disk moving frame 2; shaking and fully mixing;
13. the centrifuge tube cassette 39 is moved from the heating oscillation plate 38 to the magnetic bead separation plate 37 by the tray moving frame 2; magnetic attraction is carried out for 2 min;
14. taking 1000 mu L of pipette tips from the first pipette tip adapter 31 by using a pipette arm 1, and sucking waste liquid from a centrifuge tube box 39;
15. taking 1000 mu L of gun heads from the first gun head adapter 31 by using a liquid transfer arm 1; sucking 600. mu.L of the reagent J from the J position of the reagent tank carrier 35; to centrifuge tube cassette 39;
16. transferring the centrifuge tube box 39 from the magnetic bead separation disc 37 to the heating oscillation disc 38 by using the disc transfer frame 2, and oscillating and mixing for 15min at room temperature;
17. the centrifuge tube box 39 is moved from the heating and shaking disk 38 to the magnetic bead separation disk 37 by the moving disk rack 2, the magnetic field is absorbed for 2min, 1000 mu L of pipette tips are taken from the centrifuge tube box 39 by the pipette arm 1 in the first pipette tip adapter 31, and waste liquid is absorbed;
18. taking 1000 mu L of gun heads from the first gun head adapter 31 by using a liquid transfer arm 1; sucking 600. mu.L of reagent K from the K position of the reagent tank carrier 35; to centrifuge tube cassette 39;
19. transferring the centrifuge tube box 39 from the magnetic bead separation disc 37 to the heating oscillation disc 38 by using the disc transfer frame 2, and oscillating and mixing for 15min at room temperature;
20. the centrifuge tube cassette 39 is transferred from the heating and shaking tray 38 to the magnetic bead separation tray 37 by the transfer tray 2, and then magnetic attraction is performed for 2 min. Taking 1000 mu L of pipette tips from the magnetic bead separation disc 37 by using a pipette arm on a third pipette tip adapter 34, and sucking waste liquid;
21. taking 1000 mu L of gun heads from the third gun head adapter 34 by using a liquid transfer arm 1; sucking 600. mu.L of reagent K from the K position of the reagent tank carrier 35; to centrifuge tube cassette 39;
22. transferring the centrifuge tube box 39 from the magnetic bead separation disc 37 to the heating oscillation disc 38 by using the disc transfer frame 2, and oscillating and mixing for 15min at room temperature;
23. the centrifuge tube cassette 39 is transferred from the heating and shaking tray 38 to the magnetic bead separation tray 37 by the transfer tray 2, and then magnetic attraction is performed for 2 min. Taking 1000 mu L of pipette tips from the third pipette tip adapter 34 by using a pipette arm, and sucking waste liquid from the centrifuge tube box 39;
24. drying at room temperature for 10 min;
25. taking 200 mu L of gun heads from the second gun head adapter 32 by using a liquid transfer arm; aspirating 40 μ L of reagent E from the reagent tank carriage 35E; to centrifuge tube cassette 39;
26. transferring the centrifuge tube box 39 from the magnetic bead separation disc 37 to the heating oscillation disc 38 by using the disc transfer frame 2, and oscillating and mixing for 10min at room temperature;
27. the centrifugal tube box 39 is moved from the heating and shaking disk 38 to the magnetic bead separation disk 37 by the moving disk frame 2, and the magnetic attraction is carried out for 2 min;
28. using a pipette arm, 200. mu.L of pipette tips were taken from the second pipette tip adapter 32, and 40. mu.L of eluates were taken from the centrifuge tube cassette 39 and stored in the last 24 wells of the PCR tube covered with a drum cover by the transfer temperature control adapter 36. The product can be subjected to molecular experiments such as PCR, RT-PCR, TMA, NASBA, 3SR, LCR, enzyme digestion reaction, ligation reaction, transformation, transfection and the like.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (9)
1. A liquid workstation applied to biology comprises a working platform (6), a working disk surface (3), a liquid transfer arm (1) and a liquid transfer rack (2), wherein the working disk surface (3) is arranged on the working platform (6), an equipment box (60) is vertically arranged on the rear top surface of the working platform (6), the liquid transfer arm (1) is arranged at the top of the equipment box (60) through an x-axis driving mechanism, a y-axis driving mechanism and a Z-axis driving mechanism, and the x-axis of the liquid transfer arm (1) is parallel to the equipment box (60);
the device is characterized in that the area of the working disk surface (3) comprises nine working disk positions which are arranged in a grid manner, wherein a first gun head adapter (31), a second gun head adapter (32), a sample collecting carrier (33), a third gun head adapter (34), a reagent groove carrier (35), a temperature control adapter (36), a magnetic bead mixer (4), a magnetic bead separating disk (37) and a heating vibration disk (38) are sequentially arranged on the working disk positions from inside to outside and from left to right;
a plurality of standard gun heads are placed on the first gun head adapter (31), the second gun head adapter (32) and the third gun head adapter (34), a suction head mechanism for sucking gun heads on the first gun head adapter (31), the second gun head adapter (32) and the third gun head adapter (34) is arranged on the liquid transfer arm (1), a standard pore plate for placing an experimental to-be-detected product is arranged on the sample collection carrier (33), a plurality of standard reagent bottles are placed on the reagent groove carrier (35), a plurality of specifications of PCR tubes are placed on the top of the temperature control adapter (36), a temperature control device for heating and refrigerating the PCR tubes is arranged at the bottom of the temperature control adapter (36), and a waste object area is arranged on the left side of the magnetic bead mixer (4);
magnetic bead separating disc (37) include bar magnet (373), the heating is shaken dish (38) and is included the spiral oscillation mechanism of bottom, and magnetic bead separating disc (37) or heating are shaken dish (38) and are put on the shelf and are equipped with and move dish frame (2), move dish frame (2) and put on the shelf and be equipped with centrifugal tube dish (39), move dish frame (2) and erect on the work dish position on work dish face (3), be equipped with on liquid-moving arm (1) and be used for getting the mechanism with the hook that moves dish frame (2) whole and lift up.
2. The liquid workstation applied to biology according to claim 1, wherein the centrifuge tube tray (39) is provided with a centrifuge tube placing area (390) for placing a centrifuge tube body and a centrifuge tube fixing area (391) for fixing a centrifuge tube cover.
3. Liquid workstation for biology according to claim 1, wherein the tray moving frame (2) comprises a solution tray support frame (22) and a tray moving hook frame (20), wherein a centrifuge tube tray (39) is erected on the solution tray support frame (22), the solution tray support frame (22) is arranged around the tray position opening of the work tray surface (3), a frame groove for supporting the solution tray support frame (22) is arranged at the edge of the tray position opening, and the tray moving hook frame (20) is vertically arranged on the side wall of the solution tray support frame (22).
4. The liquid working station applied to biology according to claim 3, wherein a lifting seat (10) moving in a vertical direction is installed at the bottom of the Z axis of the liquid transferring arm (1), the hooking mechanism comprises a hook head block (11) installed on the lifting seat (10) and a hook hole block (21) installed on the side wall of the top of the tray transferring hook frame (20), a plurality of hook nails are vertically fixed on the top surface of the hook head block (11), and a plurality of nail holes for inserting the hook nails are opened on the bottom surface of the hook block (21).
5. The liquid workstation applied to biology according to claim 1, wherein the waste area comprises a waste liquid tank (301), a waste suction head removing device (302) and a gun head recovery box (303), the gun head recovery box (303) is arranged at the bottom of the working plate surface (3) and is positioned below the waste suction head removing device (302), the suction head device comprises a gun head plugging rod (12) arranged at the bottom of a Z shaft of the liquid transferring arm (1), the waste suction head removing device (302) comprises a suction head frame, and an opening hole for clamping the gun head plugging rod (12) is formed in a top plate of the suction head frame.
6. The liquid workstation applied to biology according to claim 5, wherein a through air channel is arranged in the gun head insertion rod (12), one end of the air channel is opened at the bottom end of the gun head insertion rod (12) and is used for generating negative pressure liquid suction for the gun head, and the other end of the air channel is communicated with a gas suction end of a liquid pump built in the equipment box (60) through an air pipe.
7. The biological fluid workstation as claimed in claim 1, wherein the magnetic rod (373) is integrally and vertically fixed on a magnetic rod base (374), the magnetic rod base (374) is fixedly installed in the working platform (6), and the magnetic rod (373) is installed to be opposite to the tube hole position of the centrifuge tube on the centrifuge tube disk (39).
8. The liquid workstation applied to biology according to claim 1, wherein the spiral oscillating mechanism comprises a motor base (382), an oscillating tube disk (381), and an oscillating motor (383), the motor base (382) is fixedly installed in the working platform (6), the oscillating motor (383) is installed at the center of the bottom surface of the motor base (382), a motor shaft of the oscillating motor (383) is horizontally provided with the oscillating tube disk (381) through an eccentric cam block (384), the top surface of the oscillating tube disk (381) is provided with a plurality of tube body grooves for placing centrifuge tubes, the centrifuge tube disk (39) is placed on the tube body groove of the oscillating tube disk (381) through a position where the moving plate disk (2) is placed corresponding to a centrifuge tube, and the bottom of the oscillating tube disk (381) is provided with a heating film.
9. The liquid workstation applied to biology according to claim 1, wherein the magnetic bead mixer (4) comprises a magnetic bead container (40), a container carrying device (41), an eccentric vibrator (42), a buffer device (43) and a mixing motor (44), the magnetic bead container (40) is sleeved in the container carrying device (41), the center of the bottom of the container carrying device (41) is in transmission connection with a motor shaft of the mixing motor (44) through the eccentric vibrator (42), and the buffer device (43) for limiting the left-right swing amplitude of the container carrying device (41) is arranged on two sides of the bottom of the container carrying device (41).
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| CN202011261784.5A CN112210476A (en) | 2020-11-12 | 2020-11-12 | Be applied to biological liquid workstation |
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| CN202011261784.5A CN112210476A (en) | 2020-11-12 | 2020-11-12 | Be applied to biological liquid workstation |
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