LU503307B1 - Device for automatically preparing ribonucleic acid and preparing method - Google Patents
Device for automatically preparing ribonucleic acid and preparing method Download PDFInfo
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- LU503307B1 LU503307B1 LU503307A LU503307A LU503307B1 LU 503307 B1 LU503307 B1 LU 503307B1 LU 503307 A LU503307 A LU 503307A LU 503307 A LU503307 A LU 503307A LU 503307 B1 LU503307 B1 LU 503307B1
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- 229920002477 rna polymer Polymers 0.000 title claims abstract description 82
- 238000000034 method Methods 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 claims abstract description 75
- 238000005119 centrifugation Methods 0.000 claims abstract description 60
- 239000007788 liquid Substances 0.000 claims abstract description 60
- 238000003860 storage Methods 0.000 claims abstract description 57
- 230000007246 mechanism Effects 0.000 claims abstract description 50
- 238000009434 installation Methods 0.000 claims abstract description 43
- 239000000463 material Substances 0.000 claims abstract description 9
- 230000032798 delamination Effects 0.000 claims description 3
- 239000000243 solution Substances 0.000 description 80
- 238000006243 chemical reaction Methods 0.000 description 25
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 20
- 239000003153 chemical reaction reagent Substances 0.000 description 18
- 239000011259 mixed solution Substances 0.000 description 17
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 11
- 230000008569 process Effects 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 238000010438 heat treatment Methods 0.000 description 6
- 238000000605 extraction Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 4
- 238000012864 cross contamination Methods 0.000 description 3
- 238000002123 RNA extraction Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000013517 stratification Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N2035/00465—Separating and mixing arrangements
- G01N2035/00495—Centrifuges
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Abstract
Device For Automatically Preparing Ribonucleic Acid And Preparing Method Disclosed are a device for automatically preparing ribonucleic acid and a preparing method. The device for automatically preparing RNA includes a centrifugation device, a solution preparation device, a suction device and a control device. The centrifugation device is provided with a centrifugation cavity, and the centrifugation cavity is provided with a feed cavity and a discharge cavity communicating with the feed cavity. The solution preparation device includes an installation seat movably provided in the solution preparation device. The installation seat is provided with a plurality of cavities, and each cavity is provided with a liquid outlet to enable the liquid outlet of each cavity to pass through and communicate with the feed cavity sequentially in a stroke of the installation seat. The suction device includes a cylinder and a suction mechanism. A liquid storage cavity is provided in the cylinder, and a pipette tip is connected to a discharge end of the cylinder to make material in the discharge cavity sucked into the liquid storage cavity by the pipette tip when the suction device is sucking. The control device is electrically connected to each of the suction device and the solution preparation device, to control the centrifugation device, the suction device and the solution preparation device to move.
Description
DEVICE FOR AUTOMATICALLY PREPARING RIBONUCLEIC | U503307
ACID AND PREPARING METHOD
[0001] This application claims priority to Chinese Patent Application No. 202210407124.6, filed on April 18, 2022, the entire disclosure of which is incorporated herein by reference.
[0002] The present disclosure relates to the technical field of ribonucleic acid (RNA) preparation and extraction, in particular to a device for automatically preparing RNA and a preparing method.
[0003] The ribonucleic acid (RNA) extraction technology is widely used in key biomedical experiments, such as the polymerase chain reaction (PCR) amplification. At present, the existing devices for automatically preparing RNA are generally based on the Trizol reagent method to extract RNA. Based on the Trizol reagent method, the process for extracting RNA includes several key steps such as adding the Trizol reagent and chloroform, mixing and stirring the mixed solution of the Trizol reagent and chloroform, centrifuging the mixed solution of the
Trizol reagent and chloroform, and sucking the supernatant. Currently, these operations are mostly performed by manual operations, and after the operations are repeated for many times, the operational accuracy is easily reduced and fatigue is easily increased, which is not conducive to scale production. In the existing devices for extracting RNA, the layered interface of the centrifuged mixed solution is identified by human eyes, and the supernatant is sucked by the pipette, resulting in a low operation accuracy and a low efficiency. In addition, RNA can be bound to proteins and DNA, and there is a little protein and DNA pollution in the external environment such as the air. Since the existing devices for extracting RNA cannot provide a sealed environment, it is difficult to extract high-purity RNA. Furthermore, the reagent and the cell solution cannot be added automatically in the existing devices for extracting RNA, but are added and extracted basically by manually controlling the volume range of the pipette.
[0004] In the existing devices for extracting RNA, by manual operations and human eyes to identify the layered interface, the reagent and the cell solution can be added and extracted. The layered interface of the centrifuged mixed solution is identified by human eyes, resulting in a low accuracy and a low efficiency. In the related art, although operations of the RNA extraction process are performed in a fully sealed environment, the automation degree is relatively low, and the reagent is added and extracted by manual operations. After centrifugation, the mixed solution will be stratified to generate the supernatant. In the existing technology, the layered interface is identified by human eyes, and the supernatant is sucked by the pipette, which makes operating errors easily caused and the workload of the operator increased. Based on the Trizol reagent 509907 method, the process for extracting RNA includes many steps, and the waiting time of each step is long. In this case, the process for extracting RNA cannot be fully automated and unattended.
[0005] The main objective of the present disclosure is to provide a device for automatically preparing ribonucleic acıd (RNA) and a preparing method, aiming to provide a device for automatically preparing RNA and a preparing method to realize the automation and intelligence of RNA preparation and extraction.
[0006] In order to achieve the above objectives, the present disclosure provides a device for automatically preparing RNA, including:
[0007] a centrifugation device provided with a centrifugation cavity, the centrifugation cavity is provided with a feed cavity and a discharge cavity communicating with the feed cavity;
[0008] a solution preparation device for adding RNA preparation solution to the feed cavity, the solution preparation device includes an installation seat movably provided in the solution preparation device, the installation seat is provided with a plurality of cavities, and each cavity is provided with a liquid outlet to enable the liquid outlet of each cavity to pass through and communicate with the feed cavity sequentially in a stroke of the installation seat;
[0009] a suction device including a cylinder and a suction mechanism, a liquid storage cavity is provided in the cylinder, and a pipette tip is detachably connected to a discharge end of the cylinder to make material in the discharge cavity sucked into the liquid storage cavity by the pipette tip when the suction device is sucking; and
[0010] a control device electrically connected to each of the centrifugation device, the suction device and the solution preparation device, to control the centrifugation device, the suction device and the solution preparation device to move.
[0011] In an embodiment, the solution preparation device further includes a turntable rotatably provided on the installation seat, the turntable is provided above the feed cavity and configured to rotate along a vertical axis, and the plurality of cavities are provided at intervals along a circumference of the turntable.
[0012] In an embodiment, the solution preparation device further includes a plurality of storage containers; a plurality of installation positions are provided at the turntable at intervals along the circumference of the turntable, and each installation position is for the storage container to be detachably installed on the installation position; and when the storage container is installed on the installation position, an inner cavity of the storage container is the cavity.
[0013] In an embodiment, the cylinder is provided above the discharge cavity, the suction mechanism includes a piston rod and a first driving mechanism, the piston rod provided in the 203907 liquid storage cavity is movable up and down, and the first driving mechanism provided outside the cylinder is connected to the piston rod, and the first driving mechanism is for driving the piston rod to move upward, to make the material in the discharge cavity enter the liquid storage cavity by the pipette tip.
[0014] In an embodiment, the device further includes a storage box, the storage box, the solution preparation device and the suction device are provided at intervals, the storage box is provided with a plurality of placement positions, and each placement position is for storing one pipette tip.
[0015] In an embodiment, the cylinder is configured to reciprocate between the placement position and the discharge cavity, the suction device further includes a second driving mechanism for driving the cylinder to move, and the second driving mechanism includes a rack and a gear engaging with the rack, and the rack installed on a side of the cylinder is configured to reciprocate between the placement position and the discharge cavity, and the gear is for driving the rack to move upward and downward to drive the cylinder to move upward and downward.
[0016] In an embodiment, the suction device further includes a connection structure provided at a discharge end of the cylinder, and the connection structure is for clamping with the pipette tip when the cylinder is close to the placement position.
[0017] In an embodiment, the suction device further includes a disassembly structure for disassembling the pipette tip from the cylinder, the disassembly structure includes a sliding block and a third driving mechanism, the sliding block provided outside the cylinder is configured to slid upward and downward, and the third driving mechanism is for driving the sliding block to slide upward to make the pipette tip clamped with the cylinder, and for driving the sliding block to slide downward to disconnect the pipette tip from the cylinder.
[0018] In an embodiment, the device further includes a light emitting element and a receiving element, both the light emitting element and the receiving element are provided at a bottom of the discharge cavity, the light emitting element is for emitting a light beam to a layered liquid surface, and the receiving element is for receiving a light beam emitted by the light emitting element and reflected by the layered liquid surface, to determine delamination of the layered liquid surface according to intensity of a received light beam.
[0019] In addition, the present disclosure further provides a preparing method, based on the device for automatically preparing RNA, including following operations:
[0020] controlling the installation seat to move, to make each cavity closed to the feed cavity sequentially;
[0021] when each cavity 1s close to the feed cavity, controlling the cavity close to the feed cavity to communicate with the feed cavity, to make the RNA preparation solution enter the feed 203907 cavity;
[0022] centrifuging the feed cavity; and
[0023] controlling the suction device to suck liquid in an upper layer of the discharge cavity.
[0024] In technical solutions of the present disclosure, the centrifugation device is provided with a feed cavity and a discharge cavity communicating with the feed cavity. The installation seat is movably provided in the solution preparation device. Through the movements of the installation seat, the liquid outlet of each cavity can pass through and communicate with the feed cavity sequentially. The control device is for controlling the installation seat to move to realize an automatic RNA preparation, thereby avoiding that the RNA preparation solution is measured and added to the feed cavity by manual operations for many times. In this way, the efficiency can be improved. The solution in the feed cavity enters the discharge cavity after the preparation is finished, and the control device is for controlling the centrifugation device to perform centrifugation. After the centrifugation is finished, the suction structure is for sucking the material in the discharge cavity into the liquid storage cavity by the pipette tip. The control device is for controlling the suction device to perform suction, to make the solution be sucked automatically into the liquid storage cavity after the preparation is finished, and then the RNA can be eventually obtained. The pipette tip is detachably connected to the cylinder, in this way, a cross-contamination of solutions caused by suction can be effectively avoided, and an additional cleaning device is not required. Therefore, not only the preparation cost of the device for automatically preparing RNA can be saved, but also the suction efficient is higher. In the present disclosure, RNA can be prepared and extracted automatically by the device for automatically preparing RNA, which not only avoids a situation that the preparation efficiency is reduced due to a manual intervention, but also prevents RNA from being polluted during the preparation and extraction process.
[0025] To illustrate the technical solutions according to the embodiments of the present disclosure or the related art more clearly, the accompanying drawings for describing the embodiments or the related art are introduced briefly in the following. Apparently, the accompanying drawings in the following description are only some embodiments of the present disclosure. Persons of ordinary skill in the art can derive other drawings from the structures of the accompanying drawings without creative efforts.
[0026] FIG. 1 is an overall schematic view of a device for automatically preparing ribonucleic acid (RNA) according to an embodiment of the present disclosure.
LU503307
[0027] FIG. 2 is a schematic structural view of a centrifugation device in FIG. 1.
[0028] FIG. 3 1s an enlarged schematic view of place À in FIG. 1.
[0029] FIG. 4 is a schematic structural view of a solution preparation device in FIG. 1. 5 [0030] FIG. 5 is a top view of the solution preparation device in FIG. 1.
[0031] FIG. 6 is a schematic structural view of a suction device and a storage box in FIG. 1.
[0032] FIG. 7 is a schematic flowchart of a method for preparing RNA according to an embodiment of the present disclosure.
[0033] Description of reference numbers:
Name Name number number device for automatically preparing a . 100 311 liquid storage cavity
RNA
. second driving 13 reaction bottle 33 . mechanism solution preparation device sliding block
[0034] The realization of the objective, functional characteristics, and advantages of the present disclosure are further described with reference to the accompanying drawings.
[0035] The technical solutions of the embodiments of the present disclosure will be described clearly in the following with reference to the accompanying drawings. It is obvious that the embodiments described are only some rather than all of the embodiments of the present disclosure. Based on the embodiments of the present disclosure, all other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of the present. 509907 disclosure.
[0036] It should be noted that all the directional indications (such as up, down, left, right, front, rear...) in the embodiments of the present disclosure are only used to explain the relative positional relationship, movement, or the like of the components in a certain posture (as shown in the drawings). If the specific posture changes, the directional indication will change accordingly.
[0037] Besides, the descriptions associated with, e.g., “first” and “second,” in the present disclosure are merely for descriptive purposes, and cannot be understood as indicating or suggesting relative importance or impliedly indicating the number of the indicated technical feature. Therefore, the feature associated with “first” or “second” can expressly or impliedly include at least one such feature. In addition, the meaning of “and/or” described in the whole disclosure includes three parallel schemes. Taking “a and/or b” as an example, it includes scheme a, scheme b, or schemes that a and b are satisfied at the same time. In addition, the technical solutions of the various embodiments can be combined with each other, but the combinations must be based on the realization of those skilled in the art. When the combination of technical solutions is contradictory or cannot be achieved, it should be considered that such a combination of technical solutions does not exist, nor does it fall within the scope of the present disclosure.
[0038] In the existing devices for extracting ribonucleic acid (RNA), by manual operations and human eyes to identify the layered interface, the reagent and the cell solution can be added and extracted. The layered interface of the centrifuged mixed solution is identified by human eyes, resulting in a low accuracy and a low efficiency. In the related art, although operations of the RNA extraction process are performed in a fully sealed environment, the automation degree 1s relatively low, and the reagent is added and extracted by manual operations. After centrifugation, the mixed solution will be stratified to generate the supernatant. In the existing technology, the layered interface is identified by human eyes, and the supernatant is sucked by the pipette, which makes operating errors easily caused and the workload of the operator increased. Based on the Trizol reagent method, the process for extracting RNA includes many steps, and the waiting time of each step is long. In this case, the process for extracting RNA cannot be fully automated and unattended. In view of this, the present disclosure provides a device for automatically preparing RNA 100 and a preparing method. As shown in FIGS. 1 to 5, in an embodiment, the present disclosure provides a device for automatically preparing RNA 100.
[0039] It should be noted that the method for preparing RNA is to extract RNA based on the
Trizol reagent method, which generally includes following steps. Step 1, adding the Trizol reagent and mixing the solution, then letting the mixed solution stand at room temperature for 203907 five minutes. Step 2, adding 200 ul of chloroform to the mixed solution and mixing the solution with chloroform gently, then letting the newly mixed solution stand at room temperature for five minutes. Step 3, centrifuging the mixed solution at 4°C and 12000pm for fifteen minutes, then sucking the layered supernatant (about 400ul) into a new microcentrifuge tube (namely a centrifuge tube). Step 4, adding 400 pl of isopropanol to the new microcentrifuge tube and mixing the solution, then letting the mixed solution stand at room temperature for ten minutes.
Step 5, centrifuging the mixed solution at 4°C and 12000rpm for ten minutes, then sucking and discarding the supernatant. Step 6, adding 1ml of pre-cooled 75% ethanol to the precipitation, then centrifuging a mixed solution of the ethanol and the precipitation at 4°C and 7500rpm for five minutes. Step 7, sucking and discarding the supernatant by a pipette, then letting the precipitate at the bottom of the centrifuge tube dry in the air for ten minutes, and after the color of the precipitation gradually changes from white to translucent, adding 11.5ul of special ribonuclease-free water to the precipitation. In this method, solutions are added and centrifuged for many times, and the supernatant is sucked for many times. Therefore, a more intelligent device is required.
[0040] As shown in FIG. 1 and FIG. 2, a device for automatically preparing RNA 100 includes a centrifugation device 1, a solution preparation device 2, a suction device 3 and a control device. The centrifugation device 1 is provided with a centrifugation cavity 131, and the centrifugation cavity 131 is provided with a feed cavity and a discharge cavity communicating with the feed cavity. The solution preparation device 2 for adding RNA preparation solution to the feed cavity, and the solution preparation device 2 includes an installation seat movably provided in the solution preparation device 2. The installation seat is provided with a plurality of cavities 21, and each cavity 21 is provided with a liquid outlet 211 to enable the liquid outlet 211 of each cavity 21 to pass through and communicate with the feed cavity sequentially in a stroke of the installation seat. The suction device 3 includes a cylinder 31 and a suction mechanism 32, and a liquid storage cavity 311 is provided in the cylinder 31. A pipette tip 5 is detachably connected to a discharge end of the cylinder 31 to make material in the discharge cavity sucked into the liquid storage cavity 311 by the pipette tip 5 when the suction device 3 is sucking. The control device is electrically connected to each of the centrifugation device 1, the suction device 3 and the solution preparation device 2, to control the centrifugation device 1, the suction device 3 and the solution preparation device 2 to move.
[0041] In the embodiment, the centrifugation device 1 is provided with a feed cavity and a discharge cavity communicating with the feed cavity. The installation seat is movably provided in the solution preparation device 2. Through the movements of the installation seat, the liquid outlet 211 of each cavity 21 can pass through and communicate with the feed cavity sequentially. 997
The control device is for controlling the installation seat to move to realize an automatic RNA preparation, thereby avoiding that the RNA preparation solution is measured and added to the feed cavity by manual operations for many times. In this way, the efficiency can be improved.
The solution in the feed cavity enters the discharge cavity after the preparation is finished, and the control device is for controlling the centrifugation device 1 to perform centrifugation. After the centrifugation is finished, the suction structure is for sucking the material in the discharge cavity into the liquid storage cavity 311 by the pipette tip 5. The control device is for controlling the suction device 3 to perform suction, to make the solution sucked automatically into the liquid storage cavity 311 after the preparation is finished, and then the RNA can be eventually obtained.
The pipette tip 5 is detachably connected to the cylinder 31, in this way, a cross-contamination of solutions caused by suction can be effectively avoided, and an additional cleaning device is not required. Therefore, not only the preparation cost of the device for automatically preparing RNA 100 can be saved, but also the suction efficient is higher. In the present disclosure, RNA can be prepared and extracted automatically by the device for automatically preparing RNA 100, which not only avoids a situation that the preparation efficiency is reduced due to a manual intervention, but also prevents RNA from being polluted during the preparation and extraction process.
[0042] It should be noted that in the embodiment, the method for preparing RNA is based on the Trizol reagent method to extract RNA. When the Trizol reagent method is used to extract
RNA, the RNA preparation solution will include toxic substances such as phenol, guanidine isothiocyanate and B-mercaptoethanol, and the volatilized toxic solutions which are likely to endanger human health. In the present disclosure, the device for automatically preparing RNA 100 can effectively protect human health.
[0043] As shown in FIG. 4 and FIG. 5, further, the solution preparation device 2 further includes a turntable 22 rotatably provided on the installation seat. The turntable 22 is provided above the feed cavity and configured to rotate along a vertical axis. The plurality of cavities 21 are provided at intervals along a circumference of the turntable 22.
[0044] In the embodiment, the turntable 22 rotatably provided on the installation seat is provided above the feed cavity. The liquid in the plurality of cavities 21 is configured to pass through the liquid outlet 211, and then enter the feed cavity due to the gravity. In this way, the structure is simple and easy to implement. The plurality of cavities 21 are provided at intervals along a circumference of the turntable 22. Therefore, when the turntable 22 rotates, the plurality of cavities 21 can sequentially pass above the feed cavity and inject RNA preparation solutions with different compositions into the feed cavity. By controlling the rotation angle of the turntable
22, the cavity 21 can communicate with the feed cavity easily, and various solutions can be added without setting a specific adding order. In some cases, if the solution added in a certain 203907 cavity 21 is not enough and needs to be added again, it only needs to rotate the turntable 22 to make the solution preparation device 2 used under various adding situations, and the solution preparation device 2 has a wider application prospect. The solution preparation device 2 is not limited to the turntable 22. In another embodiment, the solution preparation device 2 may include a rotating shaft instead of the turntable 22, and the plurality of cavities 21 are rotatably installed on the rotating shaft through connecting rods. When the rotating shaft rotates, the plurality of cavities 21 are driven to rotate. The specific form of the solution preparation device 2 is not limited in the present disclosure, as long as the plurality of cavities 21 communicate with the feed cavity sequentially.
[0045] Furthermore, as shown in FIG. 5, the solution preparation device 2 further includes a plurality of storage containers. A plurality of installation positions 221 are provided at the turntable 22 at intervals along the circumference of the turntable 22, and each installation position 221 is for the storage container to be detachably installed on the installation position 221.
When the storage container is installed on the installation position 221, an inner cavity of the storage container is the cavity 21.
[0046] In the embodiment, a plurality of installation positions 221 are provided at the turntable 22 at intervals along the circumference of the turntable 22, which enables the storage container to be detachably installed on the installation positions 221. In this way, the storage container can be quickly replaced, and when RNA are prepared in mass production, the preparing time can be saved and efficiency can be improved. In the present disclosure, the installation method between the storage container and the turntable 22 is not specifically limited, as long as the storage container can be detachably installed on the turntable 22. The storage container can be clamped or bound on the turntable 22. The storage container is further provided a piston rod 321 and a driving mechanism for driving the gear and the rack. The piston rod 321 provided in a cavity 21 of the storage container is movable up and down, and the piston rod 321 is connected to the rack. The gear engaging with the rack is for driving the rack to move upward and downward to drive the piston rod 321 to move upward and downward. When the piston rod 321 moves downward, the liquid in the cavity 21 of the storage container is discharged to the feed cavity through the liquid outlet 211. The piston rod 321 is driven by the rack and the gear, which not only enables the liquid volume added to the feed cavity to be controlled accurately and conveniently, but also makes the liquid in the storage container replenished conveniently when the liquid in the storage container is used up. In this embodiment, since RNA is extracted based the Trizol reagent method, the installation seat is provided with six cavities, and the six cavities are respectively for trizol, chloroform, isopropanol, 75% ethanol, ribonuclease-free water and clear water to be added. Correspondingly, the cavities are the Trizol cavity, the chloroform cv 203907 the isopropanol cavity, the ethanol cavity, the ribonuclease-free water cavity and the clear water cavity. In another embodiment, if different preparation method is applied in an embodiment, or more or less solution is used in an embodiment, adaptive adjustments can be made in the embodiments.
[0047] As shown in FIG. 5, the cylinder 31 is provided above the discharge cavity. The suction mechanism 32 includes a piston rod 321 and a first driving mechanism 322, and the piston rod 321 provided in the liquid storage cavity 311 is movable up and down. The first driving mechanism 322 provided outside the cylinder 31 is connected to the piston rod 321, and the driving mechanism is for driving the piston rod 321 to move upward, to make the material in the discharge cavity enter the liquid storage cavity 311 by the pipette tip 5. In this embodiment, the first driving mechanism 322 is for driving the piston rod 321 to move upward and downward to perform suction. The first driving mechanism 322 includes a rack and a gear, and the rack connected to the first piston rod 321 is movable up and down. The gear engaging with the rack rotates to drive the rack to move upward and downward, which makes the piston rod 321 move upward and downward in the liquid storage cavity 311 to perform suction. In this way, the suction mechanism 32 can better control the volume of the sucked liquid when sucking, to avoid an excessive suction.
[0048] Furthermore, as shown in FIG. 5, the device for automatically preparing RNA 100 further includes a storage box 4. The storage box 4, the solution preparation device 2 and the suction device 3 are provided at intervals. The storage box 4 is provided with a plurality of placement positions 41, and each placement position 41 is for storing one pipette tip 5.
[0049] In this embodiment, when RNA is prepared, to avoid cross-contamination of the liquid on the pipette tip 5, the pipette tip 5 will be discarded after the supernatant is sucked, and then the pipette tip 5 will be replaced with a new pipette tip 5 for continue suction. Thus, a storage box 4 is provided for storing the discarded pipette tip 5 and replacing a new pipette tip 5.
In this way, not only the step of replacing the pipette tips 5 by manual work can be reduced, and manpower can be reduced to realize the intelligence, but also the pollution from the outside world can be reduced. In this embodiment, the storage box 4, the solution preparation device 2 and the suction device 3 are provided at intervals, which is equivalent to provide three working stations in the device for automatically preparing RNA 100. The solution preparation device 2 is provided at the first working station, and the storage box 4 is provided at the second working station, and the suction device 3 is provided at the third working station. The RNA solution is prepared at the first working station by the centrifugation cavity 131, and the pipette tip 5 is assembled at the second working station by the suction device 3, and a suction is performed at the third working station by the suction device 3. In this way, by a reasonable division of labor 99997 the three working stations can form a whole set of processes. Therefore, the RNA preparation efficiency can be improved, and the RNA preparation can be more suitable in a large scale.
[0050] Furthermore, the cylinder 31 is configured to reciprocate between the placement position 41 and the discharge cavity. The suction device 3 further includes a second driving mechanism 33 for driving the cylinder 31 to move, and the second driving mechanism 33 includes a rack and a gear engaging with the rack. The rack installed on a side of the cylinder 31 is configured to reciprocate between the placement position 41 and the discharge cavity, and the gear is for driving the rack to move upward and downward to drive the cylinder 31 to move upward and downward.
[0051] In this embodiment, the cylinder 31 is configured to reciprocate between the placement position 41 and the discharge cavity, which is convenient for assembling the pipette tip 5, disassembling the pipette tip 5 and performing suction. In the three steps of assembling the pipette tip 5, disassembling the pipette tip 5 and performing suction, the cylinder 31 needs to move upward and downward for assembling the pipette tip 5, disassembling the pipette tip 5 and performing better suction. Therefore, the suction device 3 is provided with a second driving mechanism 33, and the driving structures of the gear and the rack are simple, which is convenient for assembling and controlling.
[0052] Furthermore, the suction device 3 further includes a connection structure provided at a discharge end of the cylinder 31, and the connection structure is for clamping with the pipette tip 5 when the cylinder 31 is close to the placement position 41.
[0053] An end of the pipette tip 5 connected to the cylinder 31 is an installation end, and the cylinder 31 is clamped with the pipette tip 5. In this embodiment, the specific clamping manner can be an interference socketing, and the diameter of the installation end of the pipette tip 5 is larger than that of the cylinder 31. The cylinder 31 can be clamped with the pipette tip 5 through an interference fit. In the embodiment, to make the connection between the pipette tip 5 and the cylinder 31 more stable, the cylinder 31 is provided with a protrusion, and the pipette tip 5 is provided with a depression or a clamping plate. By a cooperation of the protrusion and the depression or the clamping plate, the connection between the pipette tip 5 and the cylinder 31 is more stable. In this way, the assembly can be easier, and the structure can be simpler.
[0054] As shown in FIG. 5, the suction device 3 further includes a disassembly structure 34 for disassembling the pipette tip 5 from the cylinder 31. The disassembly structure 34 includes a sliding block 341 and a third driving mechanism 342. The sliding block 341 provided outside the cylinder 31 is configured to slid upward and downward. The third driving mechanism 342 is for driving the sliding block 341 to slide upward to make the pipette tip 5 clamped with the cylinder 31, and for driving the sliding block 341 to slide downward to disconnect the pipette tip 5 from 9907 the cylinder 31.
[0055] In this embodiment, the pipette tip 5 needs to be assembled and disassembled frequently during the RNA preparation process. To save manpower and achieve higher automation, the suction device 3 further includes a disassembly structure 34 and the disassembly structure 34 includes a sliding block 341 and a third driving mechanism 342. The sliding block 341 is configured to slid on an outer wall of the cylinder 31. When the pipette tip 5 is assembled, the sliding block 341 moves upward, and when the pipette tip 5 is disassembled, the sliding block 341 moves downward to abut against the pipette tip 5. Since the pipette tip 5 is clamped with the cylinder 31, when the pipette tip 5 is disassembled, only a little external force is required to disconnect the pipette tip 5 from the cylinder 31. Due to gravity and the driving force of the third driving mechanism 342, the pipette tip 5 can be easily disassembled from the cylinder 31 by the sliding block 341. The number of the sliding block 341 can be set to one or more, which can be adjusted according to the actual situation and is not specifically limited here.
In this embodiment, the third driving mechanism 342 includes a crank-link mechanism. In another embodiment, the crank-link mechanism can be replaced with any mechanism, as long as the sliding block 341 can move upward and downward. For example, the third driving mechanism 342 can be a rack and gear driving mechanism, which is not specifically limited here.
[0056] As shown in FIG. 1 and FIG. 2, the centrifugation device 1 includes a reaction box 11, a driving device 12 and a reaction bottle 13. A reaction cavity is provided in the reaction box 11, and the reaction bottle 13 is rotatably provided in the reaction cavity along a vertical axis of the reaction box 11. A centrifugation cavity 131 is provided in the reaction bottle 13. The driving device 12 is for driving the reaction bottle 13 to rotate, to drive the liquid in the centrifugation cavity 131 to perform a centrifugal movement for stratification.
[0057] It should be noted that, in this embodiment, a plurality of reaction bottles 13 are provided in the reaction cavity, and each reaction bottle 13 includes a bottle body and a bottle cap. The bottle cap is for sealing the bottle body. The centrifugation device 1 further includes a link mechanism and a driving motor, and each bottle cap is connected to the driving motor by the link mechanism. The driving motor is for driving the link mechanism to move, to make each bottle cap move to open or close the bottle body. The reaction cavity is in a cylinder 31 shape, and the reaction cavity is oppositely provided with two reaction bottles 13 along a radial direction of the reaction cavity. When the centrifugation device 1 is performing centrifugation, the two reaction bottles 13 provided oppositely can make objects in the centrifugation device 1 more stable. In the present disclosure, the number of the reaction bottles 13 in the reaction cavity is an even number and the reaction bottles 13 are symmetrically provided in the reaction cavity, to improve the stability of objects in the centrifugation cavity 131. In addition, in this 203907 embodiment, the bottle cap is for sealing the bottle body, and the driving motor and the link mechanism are for driving the bottle cap to seal the bottle body. With the bottle cap, the driving motor and the link mechanism, the bottle body is sealed. In another embodiment, a detachable sealing bag or the like may also be used for sealing, and the method of sealing the reaction bottle 13 is not specifically limited here.
[0058] As shown in FIGS. 1 to 3, the device for automatically preparing RNA 100 further includes a light emitting element 6 and a receiving element 7. Both the light emitting element 6 and the receiving element 7 are provided at the bottom of the discharge cavity. The light emitting element 6 is for emitting a light beam to a layered liquid surface of the centrifugation cavity 131.
The receiving element 7 is for receiving a light beam emitted by the light emitting element 6 and reflected by the layered liquid surface, to determine delamination of the layered liquid surface according to intensity of a received light beam.
[0059] The light emitting element 6 may be a light emitting crystal or other elements capable of emitting light, and the receiving element 7 may be a receiving crystal or other elements capable of receiving, which are not limited here. According to principles of the light reflection and the light refraction generated by a light beam at the interface of two different media, the receiving element 7 is for detecting a light intensity difference between light beams emitted by the light emitting element 6 and reflected from the liquid surface, to determine whether the preset conditions are met. The specific principle is that the light emitting element 6 continuously emit light beams to the liquid surface in the discharge cavity, and at this time, the intensity of a reflected light signal received by the receiving element 7 is Q1. After the upper liquid is sucked by the suction device 3 completely, the intensity of a reflected light signal received by the receiving element 7 is Q2. When the intensity of the received signal changes from Q1 to Q2, it indicates that the suction is finished, and the control device controls the suction device 3 to move out of the discharge cavity. The intensity of the light signal is used to determine whether the layered liquid surface is sucked completely, and the structure is simple, and the layered liquid surface can be determined conveniently and quickly. In this way, not only the error due to manual identification can be avoided, but also the automation of the device for automatically preparing RNA can be improved. Furthermore, this identification method based on the intensity of the light signal can further be detected in a sealed environment, thereby avoiding substance contamination in the centrifugation cavity 131 when the layered interface is identified by human eyes.
[0060] The device for automatically preparing RNA 100 further includes a heating device.
The heating device provided outside the reaction bottle 13 is for providing a dry and sterile environment. The heating device may be a resistive heater, a pipe heater or an infrared heater 999307 etc., and the specific form of the heating device is not limited in the present disclosure.
[0061] The present disclosure further provides a preparing method, based on the device for automatically preparing RNA 100. The preparing method includes following operations.
[0062] S10, controlling the installation seat to move, to make each cavity closed to the feed cavity sequentially.
[0063] S20, when each cavity is close to the feed cavity, controlling the cavity close to the feed cavity to communicate with the feed cavity, to make the RNA preparation solution enter the feed cavity.
[0064] S30, centrifuging the feed cavity.
[0065] S40, controlling the suction device to suck liquid in an upper layer of the discharge cavity.
[0066] In this embodiment, the Trizol reagent method is used to extract RNA, and before the operation 40, the operations 20 to 30 are repeated for several times according to the actual situation.
[0067] Based on the device for automatically preparing RNA 100 and the RNA preparing method, a specific process of an embodiment for preparing RNA are described in the following.
First of all, starting heating the resistive heater to provide a sterile environment with high temperature and high pressure in the reaction box 11. Adding the cell solution to the centrifugation cavity 131 by manual work (in this embodiment, the feed cavity and the discharge cavity are the same cavity and from a centrifugation cavity 131 jointly). Controlling the Trizol cavity in the solution preparation device 2 to add the Trizol solution to the centrifugation cavity 131, and at this time, the cylinder 31 moves to the storage box 4 to assemble the pipette tip 5.
After the Trizol solution is added, the centrifugation device 1 rotates at a low speed (500 rpm/min) for 5 minutes to mix the solution in the centrifugation cavity 131. Then after letting the mixed solution stand at room temperature for 5 minutes, controlling the chloroform cavity in the solution preparation device 2 to add chloroform to the centrifugation cavity 131, and the volume of the added chloroform solution is one-fifth of the volume of the Trizol solution. In this case, the centrifugation device 1 rotates at a low speed (100 rpm/min) for 5 minutes to mix the solution in the centrifugation cavity 131, then letting the mixed solution stand at room temperature for 5 minutes. After that, the solution in the centrifugation cavity 131 is centrifuged and layered by the centrifugal device, then controlling the suction device 3 that has been assembled with the pipette tip 5 to suck the upper layer liquid out. After that, controlling the isopropanol cavity in the solution preparation device 2 to add isopropanol to the centrifugation cavity 131, and the volume of the added isopropanol solution is one-fourth of the volume of the
Trizol solution. Then the centrifugation device 1 rotates at a low speed (1000 rpm/min) for 5 203907 minutes to mix the solution in the centrifugation cavity 131, then letting the mixed solution stand at room temperature for 10 minutes. In this case, controlling the suction device 3 to be above the storage box 4, then disassembling the pipette tip 5 by the disassembly mechanism, and reassembling a pipette tip 5 by the disassembly mechanism. After the isopropanol is added, the solution in the centrifugation cavity 131 1s centrifuged and layered by the centrifugal device, then controlling the suction device 3 that has been assembled with the pipette tip 5 to suck the upper layer liquid out. After that, controlling the ethanol cavity in the solution preparation device 2 to add pre-cooled 75% ethanol to the centrifugation cavity 131, then the solution in the centrifugation cavity 131 is centrifuged and layered by the centrifugal device. Controlling the suction device 3 that has been assembled with the pipette tip 5 to suck the upper layer liquid out, and using the heating device to heat. After that, controlling the ribonuclease-free water cavity in the solution preparation device 2 to add ribonuclease-free water to the centrifugation cavity 131, and mix the solution. During mixing and centrifugating, the centrifugation cavity 131 is in a sealed state.
[0068] The above are only some embodiments of the present disclosure, and do not limit the scope of the present disclosure thereto. Under the concept of the present disclosure, any equivalent mechanism transformation made according to the description and drawings of the present disclosure, or direct/indirect application in other related technical fields fall within the scope of the present disclosure.
Claims (10)
1. À device for automatically preparing ribonucleic acid (RNA), comprising: a centrifugation device provided with a centrifugation cavity, wherein the centrifugation cavity 1s provided with a feed cavity and a discharge cavity communicating with the feed cavity; a solution preparation device for adding RNA preparation solution to the feed cavity, wherein the solution preparation device comprises an installation seat movably provided in the solution preparation device, the installation seat is provided with a plurality of cavities, and each cavity is provided with a liquid outlet to enable the liquid outlet of each cavity to pass through and communicate with the feed cavity sequentially in a stroke of the installation seat; a suction device comprising a cylinder and a suction mechanism, wherein a liquid storage cavity is provided in the cylinder, and a pipette tip is detachably connected to a discharge end of the cylinder to make material in the discharge cavity sucked into the liquid storage cavity by the pipette tip when the suction device is sucking; and a control device electrically connected to each of the centrifugation device, the suction device and the solution preparation device, to control the centrifugation device, the suction device and the solution preparation device to move.
2. The device of claim 1, wherein the solution preparation device further comprises a turntable rotatably provided on the installation seat, the turntable is provided above the feed cavity and configured to rotate along a vertical axis, and the plurality of cavities are provided at intervals along a circumference of the turntable.
3. The device of claim 2, wherein: the solution preparation device further comprises a plurality of storage containers; a plurality of installation positions are provided at the turntable at intervals along the circumference of the turntable, and each installation position is for the storage container to be detachably installed on the installation position; and when the storage container is installed on the installation position, an inner cavity of the storage container is the cavity.
4. The device of claim 1, wherein: the cylinder is provided above the discharge cavity, the suction mechanism comprises a piston rod and a first driving mechanism, the piston rod provided in the liquid storage cavity is movable up and down, and the first driving mechanism is provided outside the cylinder and connected to the piston rod, and the first driving mechanism is for driving the piston rod to move upward, to make the 203907 material in the discharge cavity enter the liquid storage cavity by the pipette tip.
5. The device of claim 1, further comprising a storage box, wherein the storage box, the solution preparation device and the suction device are provided at intervals, the storage box 1s provided with a plurality of placement positions, and each placement position 1s for storing one pipette tip.
6. The device of claim 5, wherein: the cylinder is configured to reciprocate between the placement position and the discharge cavity, the suction device further comprises a second driving mechanism for driving the cylinder to move, and the second driving mechanism comprises a rack and a gear engaging with the rack, and the rack installed on a side of the cylinder is configured to reciprocate between the placement position and the discharge cavity, and the gear is for driving the rack to move upward and downward to drive the cylinder to move upward and downward.
7. The device of claim 5, wherein the suction device further comprises a connection structure provided at a discharge end of the cylinder, and the connection structure is for clamping with the pipette tip when the cylinder is close to the placement position.
8. The device of claim 7, wherein: the suction device further comprises a disassembly structure for disassembling the pipette tip from the cylinder, the disassembly structure comprises a sliding block and a third driving mechanism, the sliding block provided outside the cylinder is configured to slid upward and downward, and the third driving mechanism is for driving the sliding block to slide upward to make the pipette tip clamped with the cylinder, and for driving the sliding block to slide downward to disconnect the pipette tip from the cylinder.
9. The device of claim 1, further comprising a light emitting element and a receiving element, wherein:
both the light emitting element and the receiving element are provided at a bottom of the discharge cavity, 1009807 the light emitting element 1s for emitting a light beam to a layered liquid surface, and the receiving element is for receiving a light beam emitted by the light emitting element and reflected by the layered liquid surface, to determine delamination of the layered liquid surface according to intensity of a received light beam.
10. A preparing method, based on the device of any one of claims 1 to 9, comprising following operations: controlling the installation seat to move, to make each cavity closed to the feed cavity sequentially; when each cavity is close to the feed cavity, controlling the cavity close to the feed cavity to communicate with the feed cavity, to make the RNA preparation solution enter the feed cavity; centrifuging the feed cavity; and controlling the suction device to suck liquid in an upper layer of the discharge cavity.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210407124.6A CN114703050A (en) | 2022-04-18 | 2022-04-18 | Automatic preparation device and preparation method of RNA |
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| LU503307A1 LU503307A1 (en) | 2023-10-30 |
| LU503307B1 true LU503307B1 (en) | 2024-02-27 |
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| CN (1) | CN114703050A (en) |
| LU (1) | LU503307B1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP3295014B2 (en) * | 1997-03-19 | 2002-06-24 | 株式会社大日本精機 | Automatic extraction device for component substances in liquid samples and automatic concentration measurement device for component substances in liquid samples |
| JP4017369B2 (en) * | 2001-09-28 | 2007-12-05 | 株式会社ヤクルト本社 | Nucleic acid extraction equipment |
| JP2007007595A (en) * | 2005-07-01 | 2007-01-18 | Institute Of Physical & Chemical Research | Apparatus and method for centrifugal concentration of solution |
| WO2009114514A2 (en) * | 2008-03-11 | 2009-09-17 | Tripath Imaging, Inc. | Integrated sequential sample preparation system |
| CN104893969A (en) * | 2013-11-29 | 2015-09-09 | 周琪 | Automatic plasmid extracting device with compact structure |
| EP3489693B1 (en) * | 2017-11-22 | 2020-09-09 | Brand Gmbh + Co Kg | Method for controlling a pipette device |
| CN207992255U (en) * | 2018-03-28 | 2018-10-19 | 苏州苏大赛尔免疫生物技术有限公司 | The pipette tips of blood sample adding device are de- to take device |
| CN110358673B (en) * | 2019-04-11 | 2022-09-13 | 武汉原生药谷生物医药科技有限公司 | Cell separation system and method |
| CN110628569A (en) * | 2019-10-16 | 2019-12-31 | 宁波美康盛德生物科技有限公司 | Full-automatic nucleic acid extraction system |
| KR102360717B1 (en) * | 2020-02-10 | 2022-02-10 | 주식회사 아이센스 | Method for auto- classifying whole blood/plasma/not-suction using reflected photo sensor |
| CN114058493A (en) * | 2020-07-30 | 2022-02-18 | 苏州创澜生物科技有限公司 | Liquid treatment device |
| CN114323837B (en) * | 2021-12-02 | 2022-10-18 | 湖北文理学院 | Automatic manufacturing device and manufacturing method for erythrocyte suspension |
| CN114703050A (en) * | 2022-04-18 | 2022-07-05 | 湖北文理学院 | Automatic preparation device and preparation method of RNA |
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| CN114703050A (en) | 2022-07-05 |
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