CN114703050A - Automatic preparation device and preparation method of RNA - Google Patents
Automatic preparation device and preparation method of RNA Download PDFInfo
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- CN114703050A CN114703050A CN202210407124.6A CN202210407124A CN114703050A CN 114703050 A CN114703050 A CN 114703050A CN 202210407124 A CN202210407124 A CN 202210407124A CN 114703050 A CN114703050 A CN 114703050A
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- 238000003860 storage Methods 0.000 claims abstract description 47
- 238000007599 discharging Methods 0.000 claims abstract description 27
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- 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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- 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
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Abstract
The invention discloses an automatic RNA preparation device and a preparation method, wherein the automatic RNA preparation device comprises a centrifugal device, a solution preparation device, a suction device and a control device, the centrifugal device forms a centrifugal cavity, the centrifugal cavity comprises a feeding cavity and a discharging cavity which are communicated, the solution preparation device comprises a movably arranged mounting seat, the mounting seat is provided with a plurality of containing cavities, each containing cavity is provided with a liquid outlet, so that in the moving stroke of the mounting seat, the liquid outlets sequentially pass through and are communicated with the feeding cavity; the suction device comprises a barrel and a suction mechanism, the barrel is in a stroke liquid storage cavity, the discharge end of the barrel is connected with the pipette head so as to suck the materials in the discharge cavity to the liquid storage cavity through the pipette head when the suction mechanism sucks the materials, and the control device is electrically connected with the suction device and the solution preparation device respectively and is used for controlling the centrifugal device, the suction device and the solution preparation device to move. The automatic RNA preparation device provided by the invention can realize automation and intellectualization of RNA preparation and extraction.
Description
Technical Field
The invention relates to the field of RNA preparation and extraction, in particular to an automatic RNA preparation device and a preparation method.
Background
The RNA extraction technology is widely applied to key biomedical experiments such as PCR amplification, and the like, and the existing RNA automatic preparation device is generally used for extracting RNA based on a Trizol reagent method. The process of extracting RNA based on the Trizol reagent method relates to a plurality of key steps of adding Trizol reagent and chloroform, uniformly mixing and stirring, centrifuging mixed liquid, sucking supernate and the like, and the operation steps are realized by adopting manual operation at present, and repeated operation for a plurality of times is easy to reduce the operation precision and increase fatigue, so that the method is not beneficial to developing batch production. The current RNA extraction device relies on human eyes to identify a mixed liquid layered interface after centrifugation, and then uses a liquid transfer gun to absorb supernatant, so that the operation precision and efficiency are low. RNA can be combined with protein and DNA mutually, trace protein pollution and DNA pollution exist in the external environment such as air, the current RNA extraction device lacks a sealed environment, high-purity RNA is difficult to extract, no automatic dosage function exists, and the dosage extraction of reagents and cell sap basically depends on the range of manually operating a pipette.
In the current RNA extraction device, the reagent and the cell sap are added and extracted by hand and human eyes basically, and the mixed solution layered interface after centrifugation is identified by the human eyes, so that the efficiency is low and the precision is poor. In the prior art, the full-sealing operation of the RNA extraction process is realized, but the automation degree is low, and the extraction and the addition of the reagent still need manual operation; the supernatant fluid of the mixed solution after centrifugation can be layered, the current technology adopts human eyes to identify a layered interface, and then a pipette is used for absorbing and discarding the supernatant fluid, so that operation errors are easy to generate, and the workload of operators is increased; the Trizol reagent method based RNA extraction has multiple operation steps, the waiting time of each step is long, and full automation and unattended operation cannot be realized.
Disclosure of Invention
The invention mainly aims to provide an automatic RNA preparation device and a preparation method, and aims to provide the automatic RNA preparation device and the preparation method, which can realize automation and intellectualization of RNA preparation and extraction.
In order to achieve the above object, the present invention provides an automatic RNA preparation apparatus, comprising:
the centrifugal device forms a centrifugal cavity which comprises a feeding cavity and a discharging cavity which are communicated;
the solution preparation device is used for adding an RNA preparation solution into the feeding cavity and comprises a movably arranged mounting seat, the mounting seat is provided with a plurality of containing cavities, each containing cavity is provided with a liquid outlet, so that the liquid outlets of the containing cavities sequentially pass through and are communicated with the feeding cavity in the moving process of the mounting seat;
the suction device comprises a barrel and a suction mechanism, the barrel forms a liquid storage cavity, the discharge end of the barrel is detachably connected with the pipette head, and when the suction mechanism sucks the materials in the discharge cavity, the materials are sucked into the liquid storage cavity through the pipette head; and the number of the first and second groups,
and the control device is respectively electrically connected with the centrifugal device, the suction device and the solution preparation device and is used for controlling the centrifugal device, the suction device and the solution preparation device to move.
Optionally, the solution preparation device further comprises a turntable, the turntable is rotatably arranged on the mounting seat, the turntable is arranged above the feeding cavity, the turntable is rotatably arranged along an up-down axis, and the plurality of cavities are arranged along the circumferential direction of the turntable at intervals.
Optionally, solution preparation facilities still includes a plurality of storage container, the carousel is equipped with a plurality of installation positions along its circumference interval, each the installation position is used for supplying storage container detachably install in the installation position, storage container install in when on the installation position, storage container's inner chamber constitutes hold the chamber.
Optionally, the barrel is arranged above the discharge chamber, the suction mechanism comprises a piston rod and a first driving mechanism, the piston rod is movably arranged in the liquid storage chamber up and down, the first driving mechanism is arranged on the outer side of the barrel and connected with the piston rod, and the driving mechanism drives the piston rod to move upwards so that the material in the discharge chamber enters the liquid storage chamber through the pipette head.
Optionally, the automatic RNA preparation apparatus further comprises a storage box, the solution preparation apparatus and the suction apparatus are respectively arranged at intervals, the storage box is provided with a plurality of placement positions, and each placement position is used for storing a pipette tip.
Optionally, the barrel is in the reciprocating movable setting between the placing position and the discharging chamber, the suction device further comprises a second driving mechanism for driving the barrel to move, the second driving mechanism comprises a gear and a rack, the rack is in the reciprocating movable setting between the placing position and the discharging chamber, the rack is mounted on the side portion of the barrel, the gear is meshed with the rack, and the gear rotates to drive the rack to move up and down so as to drive the barrel to move up and down.
Optionally, the suction device further comprises a connecting structure, the connecting structure is arranged at the discharge end of the barrel, and the connecting structure is used for being clamped with the pipette tip when the barrel is close to the placing position.
Optionally, the suction device further comprises a detaching structure for detaching the pipette tip from the barrel;
the disassembling structure comprises a sliding stop block and a third driving mechanism, the sliding stop block is arranged on the outer side of the barrel body in a vertically sliding mode, and the third driving mechanism is used for driving the sliding stop block to slide upwards so as to enable the pipetting gun head to be clamped with the barrel body and driving the sliding stop block to slide downwards so as to enable the pipetting gun head to be disconnected with the barrel body.
Optionally, the automatic RNA preparation apparatus further includes a light emitting element and a receiving element, both the light emitting element and the receiving element are disposed at the bottom of the discharge chamber, the light emitting element is configured to emit a light beam to the layered liquid surface in the centrifugal chamber, and the receiving element is configured to receive the light beam emitted by the light emitting element reflected by the layered liquid surface, so as to determine the layering condition of the layered liquid surface according to the intensity of the received light beam.
Correspondingly, the invention also provides a preparation method based on the automatic RNA preparation device, which comprises the following steps:
controlling the mounting seat to move so that the containing cavities sequentially and orderly approach the feeding cavity;
when each cavity is close to the feeding chamber, controlling the cavity close to the feeding chamber to be communicated with the feeding chamber so that the RNA preparation solution enters the feeding chamber;
centrifuging the feed chamber;
and controlling a suction device to suck the upper liquid level of the discharge chamber.
In the technical scheme of the invention, the centrifugal device comprises the feeding chamber and the discharging chamber which are communicated, the mounting seat can be movably arranged, the liquid outlets of the plurality of cavities sequentially pass through and are communicated with the feeding chamber through the movement of the mounting seat, the control device controls the movement of the mounting seat to realize the automatic preparation of RNA, the RNA preparation solution is prevented from being manually measured for multiple times and added into the feeding chamber, and the efficiency can be improved. The solution in the feeding chamber enters the discharging chamber after being prepared, the control device controls the centrifugal device to carry out centrifugation, after the centrifugation is finished, the suction structure is used for sucking the material in the discharging chamber into the liquid storage chamber through the pipette head, the control device controls the suction device to suck, the solution can be automatically sucked out after the preparation is finished, and finally the RNA is obtained. The pipette tip is detachably connected with the barrel, so that the cross contamination of solution caused by suction can be effectively avoided, a cleaning device is not required to be additionally arranged, the preparation cost of the automatic RNA preparation device is saved, and the suction efficiency is higher. The automatic RNA preparation device provided by the invention can realize automatic preparation and extraction of RNA, avoid manual participation to reduce preparation efficiency, and also can prevent RNA from being polluted in the preparation and extraction processes.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the structures shown in the drawings without creative efforts.
FIG. 1 is a schematic overall view of an embodiment of an apparatus for automatically preparing RNA according to the present invention;
FIG. 2 is a schematic diagram of the centrifuge of FIG. 1;
FIG. 3 is an enlarged schematic view at A in FIG. 1;
FIG. 4 is a schematic view of the structure of the solution preparing apparatus of FIG. 1;
FIG. 5 is a top view of the solution preparation device of FIG. 1;
FIG. 6 is a schematic view of the structure of the suction device and the storage box of FIG. 1;
FIG. 7 is a schematic flow chart of an embodiment of the RNA preparation method provided by the present invention.
The reference numbers illustrate:
the implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
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.
It should be noted that, if directional indications (such as up, down, left, right, front, and back … …) are involved in the embodiment of the present invention, the directional indications are only used to explain the relative positional relationship between the components, the movement situation, and the like in a specific posture (as shown in the drawing), and if the specific posture is changed, the directional indications are changed accordingly.
In addition, if there is a description of "first", "second", etc. in an embodiment of the present invention, the description of "first", "second", etc. is for descriptive purposes only and is not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In addition, the meaning of "and/or" appearing throughout includes three juxtapositions, exemplified by "A and/or B", including either A or B or both A and B. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention.
In the current RNA extraction device, the reagent and the cell sap are added and extracted by hand and human eyes basically, and the mixed solution layered interface after centrifugation is identified by the human eyes, so that the efficiency is low and the precision is poor. In the prior art, the full-sealing operation of the RNA extraction process is realized, but the automation degree is low, and the extraction and the addition of the reagent still need manual operation; the supernatant fluid of the mixed solution after centrifugation can be layered, the current technology adopts human eyes to identify a layered interface, and then a pipette is used for absorbing and discarding the supernatant fluid, so that operation errors are easy to generate, and the workload of operators is increased; the Trizol reagent method based RNA extraction has multiple operation steps, the waiting time of each step is long, and full automation and unattended operation cannot be realized. In view of the above, the present invention provides an automatic RNA preparation apparatus and a preparation method thereof, please refer to fig. 1 to 5, which are embodiments of the automatic RNA preparation apparatus provided herein.
It should be noted that the method for preparing RNA, which is based on Trizol reagent, generally comprises the following steps: the method comprises the following steps: adding a solution reagent Trizol, uniformly mixing, and standing at room temperature for 5 min; step two: adding 200 μ L chloroform, mixing, and standing at room temperature for 5 min; step three: the mixture was centrifuged at 12000pm for 15min at 4 ℃ and the separated supernatant was pipetted (about 400. mu.L) into a fresh Ep tube (i.e.centrifuge tube); step IV: adding 400 μ L isopropanol, mixing, and standing at room temperature for 10 min; step five: centrifuging the mixed solution at 4 ℃ and 12000rpm for 10min, and sucking and removing the supernatant by using a pipette; step (c): adding 1mL of precooled 75% ethanol into the precipitate, and centrifuging at 4 ℃ and 7500rpm for 5 min; step (c): the supernatant was aspirated off with a pipette, the pellet at the bottom of the tube was dried in air for 10min, the pellet gradually changed from white to translucent, and 11.5. mu.L of a special RNase-free water was added. This method involves multiple additions of solution and centrifugation and extraction of supernatant, and therefore a more intelligent device is needed.
Referring to fig. 1 and fig. 2, an automatic RNA preparation apparatus 100 includes a centrifugal apparatus 1, a solution preparation apparatus 2, a suction apparatus 3, and a control apparatus, wherein the centrifugal apparatus 1 forms a centrifugal chamber 131, and the centrifugal chamber 131 includes a feeding chamber and a discharging chamber which are communicated with each other; the solution preparation device 2 is used for adding an RNA preparation solution into the feeding cavity, the solution preparation device 2 comprises a movably arranged mounting seat, the mounting seat is provided with a plurality of cavities 21, each cavity 21 is provided with a liquid outlet 211, so that in the moving stroke of the mounting seat, the liquid outlets 211 of the cavities 21 sequentially pass through and are communicated with the feeding cavity; the suction device 3 comprises a cylinder 31 and a suction mechanism 32, the cylinder 31 forms a liquid storage cavity 311, the discharge end of the cylinder 31 is detachably connected with the pipette tip 5, so that when the suction mechanism 32 sucks, materials in the discharge cavity are sucked into the liquid storage cavity 311 through the pipette tip 5; the control device is electrically connected with the centrifugal device 1, the suction device 3 and the solution preparation device 2 respectively, and is used for controlling the centrifugal device 1, the suction device 3 and the solution preparation device 2 to move.
In this embodiment, the centrifugal device 1 includes the feeding chamber and the discharging chamber that are linked together, the mount pad is movably arranged, through the activity of the mount pad so that the liquid outlets 211 of the plurality of cavities 21 pass through and are linked together in sequence the feeding chamber, the control device controls the motion of the mount pad so as to realize the automatic preparation of RNA, avoid manually measuring RNA preparation solution many times and add into the feeding chamber, and can improve efficiency. The solution in the feeding chamber enters the discharging chamber after being prepared, the control device controls the centrifugal device 1 to carry out centrifugation, after the centrifugation is finished, the suction structure is used for sucking the material in the discharging chamber into the liquid storage cavity 311 through the pipette tip 5, the control device controls the suction device 3 to suck, the solution can be automatically sucked out after the preparation is finished, and finally the RNA is obtained. The pipette tip 5 is detachably connected with the barrel 31, so that cross contamination of solution caused by suction can be effectively avoided, an additional cleaning device is not required, the preparation cost of the automatic RNA preparation device 100 is saved, and the suction efficiency is higher. The automatic RNA preparation device 100 provided by the invention can realize automatic preparation and extraction of RNA, avoid manual participation to reduce preparation efficiency, and prevent pollution to RNA in the preparation and extraction processes.
It is worth to say that, in this embodiment, the method for preparing RNA selects and uses the method for extracting RNA based on the Trizol reagent method, and when the method for extracting RNA is used, the RNA preparation solution will contain toxic substances such as phenol, guanidine isothiocyanate and β -mercaptoethanol, and the volatile toxic solution is easy to harm the health, so the automatic RNA preparation apparatus 100 designed in this application can also effectively protect the health of human body.
Referring to fig. 4 and 5, further, the solution preparation apparatus 2 further includes a rotating disc 22, the rotating disc 22 is rotatably disposed on the mounting seat, the rotating disc 22 is disposed above the feeding chamber, the rotating disc 22 is rotatably disposed along an up-down axis, and the plurality of cavities 21 are disposed at intervals along a circumferential direction of the rotating disc 22.
In this embodiment, the rotating disc 22 is rotatably installed on the installation base, the rotating disc 22 is disposed above the feeding chamber, and the liquid in the cavities 21 enters the feeding chamber through the liquid outlet 211 under the action of gravity, so that the structure is simple and easy to implement. The plurality of cavities 21 are arranged at intervals along the circumferential direction of the rotating disc 22, so that when the rotating disc 22 rotates, the plurality of cavities 21 can sequentially pass through the upper part of the feeding chamber, and RNA preparation solutions with different components are injected into the feeding chamber. Through controlling the rotation angle of the turntable 22, the containing cavity 21 can be simply communicated with the feeding cavity, the adding conditions of various solutions can be met, a specific adding sequence is not required to be set, and in some conditions, if the adding amount of the solution in one of the containing cavities 21 is not enough and needs to be added again, the turntable 22 only needs to be rotated, so that the solution preparation device 2 can use various adding conditions, and has a wider application prospect. The solution preparing apparatus 2 is not limited to the turntable 22, and in another embodiment, it may be a rotating shaft, and the plurality of cavities 21 are rotatably mounted on the rotating shaft through a connecting rod, and the rotating shaft drives the plurality of cavities 21 to rotate when rotating. The specific form of the solution preparation device 2 is not limited in the present invention, and the plurality of cavities 21 may be sequentially communicated with the feeding chamber.
Further, referring to fig. 5, the solution preparation apparatus 2 further includes a plurality of storage containers, the rotary table 22 is provided with a plurality of mounting positions 221 at intervals along the circumferential direction thereof, each mounting position 221 is used for detachably mounting the storage container on the mounting position 221, and when the storage container is mounted on the mounting position 221, an inner cavity of the storage container forms the receiving cavity 21.
In this embodiment, the rotary table 22 is provided with a plurality of mounting positions 221 along the circumferential direction at intervals, so that the storage container can be detachably mounted on the mounting positions 221, and thus, the storage container can be replaced quickly, time can be saved when RNA is prepared in large quantities, and efficiency can be improved. The installation mode of the discharging container on the rotary disc 22 is not particularly limited, the discharging container can be detached, and can be clamped or bound on the rotary disc 22. Still be equipped with piston rod 321 and rack and pinion actuating mechanism on the storage container, piston rod 321 movably install from top to bottom in the inner chamber of storage container, piston rod 321 is connected with the rack, and the gear meshes with the rack, and gear rotation drives the rack and reciprocates in order to drive piston rod 321 and reciprocate. When the piston rod 321 moves downwards, the liquid in the inner cavity of the storage container is discharged to the feeding cavity through the liquid outlet 211. The driving mode of the gear rack driving piston rod 321 can accurately control the liquid capacity of the feeding cavity, so that the control is convenient, and the liquid in the storage container can be more conveniently supplemented after being used. In this embodiment, RNA is extracted according to the Trizol reagent method, so that six chambers 21 are provided for adding Trizol, chloroform, isopropanol, 75% ethanol, rnase-free water, and clean water, respectively, and the chambers correspond to a Trizol chamber, a chloroform chamber, an isopropanol chamber, an ethanol chamber, a rnase-free water chamber, and a clean water chamber, respectively. In other embodiments, adaptation may be made if there are different methods of preparation or if more or less liquid is used.
Referring to fig. 5, the cylinder 31 is disposed above the discharging chamber, the pumping mechanism 32 includes a piston rod 321 and a first driving mechanism 322, the piston rod 321 is disposed in the liquid storage chamber 311 in a vertically movable manner, the first driving mechanism 322 is disposed outside the cylinder 31 and connected to the piston rod 321, and the driving mechanism drives the piston rod 321 to move upward so that the material in the discharging chamber enters the liquid storage chamber 311 through the pipette head 5.
In this embodiment, the first driving mechanism 322 drives the piston rod 321 to move up and down for pumping, and the first driving mechanism 322 includes a rack-and-pinion structure, where the rack is movably connected to the first piston rod 321 up and down, the gear is engaged with the rack, and the gear rotates to drive the rack to move up and down, so that the piston rod 321 moves up and down in the reservoir 311 for pumping. The arrangement is such that the suction mechanism 32 can better control the amount of liquid sucked during suction, and excessive suction is avoided.
Further, referring to fig. 5, the automatic RNA preparation apparatus 100 further includes a storage box 4, the storage box 4 is separately disposed from the solution preparation apparatus 2 and the suction apparatus 3, the storage box 4 is provided with a plurality of placement positions 41, and each placement position 41 is used for storing one pipette tip 5.
In this embodiment, when preparing RNA, in order to avoid the cross contamination of the liquid on the pipette tip, the pipette tip is discarded after the supernatant is sucked, and then the pipette tip is replaced with a new one to continue the suction, so that a storage box 4 is provided for storing the discarded pipette tip 5 and replacing the new pipette tip, and the arrangement can reduce the manual replacement of the pipette tip 5, reduce manpower, realize higher intelligence, and reduce external pollution. In this embodiment, the storage box 4, the solution preparation device 2, and the pumping device 3 are respectively disposed at intervals, which is equivalent to three stations, the solution preparation device 2 is disposed at a first station, the storage box 4 is disposed at a second station, the pumping device 3 is disposed at a third station, the centrifugal chamber 131 prepares an RNA solution at the first station, the pumping device 3 is now equipped with a pipette tip 5 at the second station, and then pumps at the third station.
Further, the cylinder 31 is arranged between the placing position 41 and the discharging chamber in a reciprocating manner, the suction device 3 further comprises a second driving mechanism 33 for driving the cylinder 31 to move, the second driving mechanism 33 comprises a gear and a rack, the rack is arranged between the placing position 41 and the discharging chamber in a reciprocating manner, the rack is mounted on the side portion of the cylinder 31, the gear is meshed with the rack, and the gear rotates to drive the rack to move up and down so as to drive the cylinder 31 to move up and down.
In this embodiment, the cylinder 31 is reciprocally movably disposed between the placing position 41 and the discharging chamber, so as to facilitate assembling the pipette tip 5, disassembling the pipette tip 5, and performing a pumping operation, and in these three operations, the cylinder 31 needs to be moved up and down to perform assembling, disassembling, and better pumping, so that the second driving mechanism 33 is provided, and the driving manner of the gear and the rack is simple in structure and convenient to install and control.
Further, the suction device 3 further comprises a connecting structure, the connecting structure is arranged at the discharge end of the barrel 31, and the connecting structure is used for being clamped with the pipette tip 5 when the barrel 31 is close to the placing position 41.
The pipette tip 5 and the one end that barrel 31 is connected are the installation end, barrel 31 and pipette tip 5 joint, in this embodiment, the concrete mode of joint is interference fit, and the diameter of pipette tip 5 installation end is greater than barrel 31, barrel 31 can be through interference fit and pipette tip 5 joint. In a preferred embodiment, in order to make the connection between the pipette tip 5 and the barrel 31 more stable, a protrusion may be provided on the barrel 31, and a recess or a clamping plate may be provided on the pipette tip 5, which cooperate to achieve a more stable mounting during mounting. The arrangement makes the assembly simpler and the structure more simplified.
Referring to fig. 5, the suction device 3 further includes a detaching structure 34, and the detaching structure 34 is used for detaching the pipette tip 5 from the barrel 31; the detaching structure 34 includes a sliding block 341 and a third driving mechanism 342, the sliding block 341 is slidably disposed on the outer side of the barrel 31, and the third driving mechanism 342 is configured to drive the sliding block 341 to slide upwards to engage the pipette tip 5 with the barrel 31, and drive the sliding block 341 to slide downwards to disconnect the pipette tip 5 from the barrel 31.
In this embodiment, the pipette tip 5 needs to be frequently assembled and disassembled in the preparation process of RNA, in order to save labor and achieve higher automation, the suction device 3 further includes a disassembling structure 34, the disassembling structure 34 includes a sliding stopper 341 and a third driving mechanism 342, the sliding stopper 341 can slide on the outer side wall of the barrel 31, when the pipette tip 5 is installed, the slider moves upwards, and when disassembled, the slider moves downwards to abut against the pipette tip 5, because the connection manner of the pipette tip 5 and the barrel 31 is clamping, when disassembled, the pipette tip 5 and the barrel 31 can be disconnected by only a little external force, the slider can simply disassemble the pipette tip 5 from the barrel 31 by virtue of gravity and the driving force of the third driving device 12, and the slider can be provided with one block or more blocks, the method is not particularly limited, and may be adjusted according to actual conditions. In this embodiment, the third driving mechanism 342 includes a crank-link mechanism, and in other embodiments, a mechanism capable of moving the sliding block 341 up and down may be used instead of the crank-link mechanism, such as a rack and pinion transmission mechanism, and the like, which is not limited herein.
Referring to fig. 1 and fig. 2, the centrifugal apparatus 1 includes a reaction box 11, a driving apparatus 12 and a reaction bottle 13, the reaction chamber is formed in the reaction box 11, the reaction bottle 13 is rotatably disposed in the reaction chamber along an upward and downward axis of the reaction box 11, a centrifugal chamber 131 is formed in the reaction bottle 13, and the driving apparatus 12 drives the reaction bottle 13 to rotate so as to drive a liquid in the centrifugal chamber 131 to perform a centrifugal motion for layering.
It should be noted that, in this embodiment, a plurality of reaction bottles 13 are disposed in the reaction chamber, each reaction bottle 13 includes a bottle body and a bottle cap, the bottle cap is used for sealing the bottle body, the centrifugal device 1 further includes a link mechanism and a driving motor, each bottle cap is connected to the driving motor through the link mechanism, and the driving motor drives the link mechanism to move, so that each bottle cap moves to open and close the bottle body. The reaction chamber is cylindrical, and the reaction chamber is intracavity edge the radial of reaction chamber is equipped with two relatively reaction bottle 13, and two reaction bottle 13 that correspond the setting can make centrifugal device 1 make the object in the centrifugal device 1 more stable when doing centrifugal motion. In the present invention, the reaction bottles 13 are provided with even number in the reaction chamber and are symmetrically arranged in the reaction chamber, which is convenient for improving the stability of the object in the centrifugal chamber 131. Meanwhile, in the present embodiment, the bottle cap is used as a means for sealing the bottle body, the driving motor and the link mechanism are used as a means for driving the bottle cap to seal the bottle body, and the bottle body is sealed by both the driving motor and the link mechanism, and in other embodiments, the bottle body may be sealed by using a detachable sealing bag, and the like, and the means for sealing the reaction bottle 13 is not particularly limited.
Referring to fig. 1 to 3, the automatic RNA preparation apparatus 100 further includes a light emitting device 6 and a receiving device 7, the light emitting device 6 and the receiving device 7 are both disposed at the bottom of the discharging chamber, the light emitting device 6 is configured to emit a light beam to the layered liquid surface in the centrifugal chamber 131, and the receiving device 7 is configured to receive the light beam reflected by the layered liquid surface and emitted by the light emitting device 6, so as to determine the layering condition of the layered liquid surface according to the intensity of the received light beam.
The light emitting element 6 and the receiving element 7 may be a light emitting crystal and a receiving crystal, or may be other elements capable of emitting and receiving light, and are not limited herein. By utilizing the reflection and refraction principles of light rays generated at the interface of two different media, the receiving element 7 senses the intensity difference of the light emitted by the light-emitting element 6 reflected by the liquid level, and judges whether the preset condition is met or not and acts. The specific principle is that the light emitting element 6 continuously emits a light beam to the liquid level in the discharging chamber, at this time, the intensity of the reflected light signal received by the receiving element 7 is Q1, after the suction device 3 completely sucks the liquid level in the upper layer, the intensity of the reflected light signal received by the receiving element 7 is Q2, when the intensity of the received signal is changed from Q1 to Q2, the completion of suction is indicated, and the control device controls the suction device 3 to move out of the discharging chamber. Whether the layered liquid level is sucked or not is judged through the light signal intensity, the structure is simple, the layered liquid level can be conveniently and quickly judged, the error of manual identification is avoided, the automation of the RNA automatic preparation device can be improved, meanwhile, the identification method utilizing the light signal intensity can also be used for detecting in a closed environment, and the pollution to substances in the centrifugal cavity 131 during manual identification is also avoided.
The automatic RNA preparation apparatus 100 further includes a heating device disposed outside the reaction vial 13 for creating a high-temperature and high-pressure drying environment and a sterilization environment, and the specific form of the heating device is not limited in the present invention, and may be a thermal resistance wire heating, a pipe heater, an infrared heating, or the like.
The invention also provides a preparation method based on the automatic RNA preparation device 100, which specifically comprises the following steps:
s10, controlling the mounting seat to move so that the cavities 21 sequentially and orderly approach the feeding cavity;
s20, when each cavity 21 is close to the feeding chamber, controlling the cavity 21 close to the feeding chamber to be communicated with the feeding chamber so that the RNA preparation solution can enter the feeding chamber;
s30, centrifuging the feeding chamber;
and S40, controlling the suction device 3 to suck the upper liquid level of the discharging chamber.
In the present embodiment, since the Trizol reagent method is selected for extracting RNA, steps S20 to S30 are repeated as many times as practical before step S40.
Based on the automatic RNA preparation apparatus 100 and the RNA preparation method provided in this embodiment, a specific procedure of an embodiment of RNA preparation is as follows: firstly, a heating resistance wire is started to form a high-temperature and high-pressure sterile environment in the reaction box 11, cell sap is manually added into the centrifugal cavity 131 (in the embodiment, the feeding cavity and the discharging cavity are the same cavity and jointly form the centrifugal cavity 131), a Trizol accommodating cavity in the solution preparation device 2 is controlled to add Trizol solution into the centrifugal cavity 131, at the moment, the barrel 31 is moved to the storage box 4 to be provided with a pipetting gun head 5, after the Trizol solution is added, the centrifugal device 1 rotates at a low rotation speed (500rpm/min) for 5min to uniformly mix the liquid in the centrifugal cavity 131, and the centrifugal device stands for 5min at room temperature. And then adding chloroform with the volume being one fifth of that of the Trizol solution into the centrifugal cavity 131 by using a chloroform containing cavity in the solution preparation device 2, rotating the centrifugal cavity 1 at a low rotation speed (100rpm/min) for 5min to uniformly mix the liquid in the centrifugal cavity 131, and standing the mixture at room temperature for 5 min. The liquid in the centrifugal cavity 131 is centrifuged and layered by using the centrifugal device 1, the suction device 3 with the pipette tip 5 assembled is controlled to suck out the upper layer of liquid, then the isopropanol containing cavity in the solution preparation device 2 is controlled to add isopropanol with one quarter volume of Trizol solution into the centrifugal cavity 131, the centrifugal device 1 rotates at a low rotation speed (1000rpm/min) for 5min to mix the liquid in the centrifugal cavity 131 uniformly, the centrifugal cavity stands at room temperature for 10min, the suction device 3 is controlled to be positioned above the storage box 4 at the moment, the pipette tip 5 is detached by the detachment mechanism, and the pipette tip 5 is reassembled. After the isopropanol is added, the centrifugal device 1 is used for carrying out centrifugal layering on the liquid in the centrifugal cavity 131, the suction device 3 with the pipette tip 5 assembled is controlled to suck out the upper layer of liquid, then the ethanol containing cavity in the solution preparation device 2 is controlled to add 75% of precooled ethanol into the centrifugal cavity 131, the centrifugal device 1 is used for carrying out centrifugal layering on the liquid in the centrifugal cavity 131, the suction device 3 with the pipette tip 5 assembled is controlled to suck out the upper layer of liquid, the heating device is used for heating, then the RNase-free water containing cavity in the solution preparation device 2 is used for adding RNase-free water into the centrifugal cavity 131, and the RNase-free water is uniformly mixed. Wherein, during the mixing and centrifugation, the centrifugal cavity 131 is in a closed state.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the contents of the present specification and the accompanying drawings, or directly/indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. An automatic RNA preparation apparatus, comprising:
the centrifugal device forms a centrifugal cavity which comprises a feeding cavity and a discharging cavity which are communicated;
the solution preparation device is used for adding an RNA preparation solution into the feeding cavity and comprises a movably arranged mounting seat, the mounting seat is provided with a plurality of containing cavities, each containing cavity is provided with a liquid outlet, so that the liquid outlets of the containing cavities sequentially pass through and are communicated with the feeding cavity in the moving process of the mounting seat;
the suction device comprises a barrel and a suction mechanism, the barrel forms a liquid storage cavity, the discharge end of the barrel is detachably connected with the pipette head, and when the suction mechanism sucks the materials in the discharge cavity, the materials are sucked into the liquid storage cavity through the pipette head; and the number of the first and second groups,
and the control device is electrically connected with the centrifugal device, the suction device and the solution preparation device respectively and used for controlling the centrifugal device, the suction device and the solution preparation device to move.
2. The automatic RNA preparation device of claim 1, further comprising a turntable rotatably disposed on the mounting base, wherein the turntable is disposed above the feeding chamber, the turntable is rotatably disposed along an up-down axis, and the plurality of cavities are disposed at intervals along a circumferential direction of the turntable.
3. The automatic RNA preparation device according to claim 2, wherein the solution preparation device further comprises a plurality of storage containers, the rotary plate is provided with a plurality of mounting positions at intervals along a circumferential direction of the rotary plate, each mounting position is used for detachably mounting the storage container at the mounting position, and when the storage container is mounted at the mounting position, an inner cavity of the storage container forms the cavity.
4. The automatic RNA preparation device of claim 1 wherein the barrel is disposed above the discharge chamber, the pumping mechanism comprises a piston rod and a first driving mechanism, the piston rod is movably disposed in the liquid storage chamber up and down, the first driving mechanism is disposed outside the barrel and connected to the piston rod, and the driving mechanism drives the piston rod to move upward to allow the material in the discharge chamber to enter the liquid storage chamber through a pipette tip.
5. The automated RNA preparation apparatus of claim 1 further comprising a storage box spaced apart from the solution preparation apparatus and the aspiration apparatus, the storage box having a plurality of placement positions, each for storing a pipette tip.
6. The automatic RNA preparing apparatus of claim 5 wherein the barrel is reciprocally movably disposed between the placing position and the discharging chamber, the pumping device further comprises a second driving mechanism for driving the barrel to move, the second driving mechanism comprises a gear and a rack, the rack is reciprocally movably disposed between the placing position and the discharging chamber, the rack is mounted on a side portion of the barrel, the gear is engaged with the rack, and the gear rotates to drive the rack to move up and down to drive the barrel to move up and down.
7. The automatic RNA preparation device of claim 5 wherein the aspiration device further comprises a connection structure at the discharge end of the barrel for snap-fitting with a pipette tip when the barrel is adjacent to the placement location.
8. The automatic RNA preparation apparatus of claim 7 wherein the aspiration device further comprises a detachment mechanism for detaching a pipette tip from the barrel;
the disassembling structure comprises a sliding stop block and a third driving mechanism, the sliding stop block is arranged on the outer side of the barrel body in a vertically sliding mode, and the third driving mechanism is used for driving the sliding stop block to slide upwards so as to enable the pipetting gun head to be clamped with the barrel body and driving the sliding stop block to slide downwards so as to enable the pipetting gun head to be disconnected with the barrel body.
9. The automatic RNA preparation device according to claim 1, further comprising a light emitting element and a receiving element, wherein the light emitting element and the receiving element are both disposed at the bottom of the discharging chamber, the light emitting element is configured to emit a light beam to the layered liquid level, and the receiving element is configured to receive the light beam emitted by the light emitting element reflected by the layered liquid level, so as to determine the layering condition of the layered liquid level according to the intensity of the received light beam.
10. A method for preparing an apparatus for automatically preparing RNA according to any one of claims 1 to 9, comprising the steps of:
controlling the mounting seat to move so that the containing cavities sequentially and orderly approach the feeding cavity;
when each cavity is close to the feeding chamber, controlling the cavity close to the feeding chamber to be communicated with the feeding chamber so that the RNA preparation solution enters the feeding chamber;
centrifuging the feed chamber;
and controlling a suction device to suck the upper liquid level of the discharge chamber.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210407124.6A CN114703050A (en) | 2022-04-18 | 2022-04-18 | Automatic preparation device and preparation method of RNA |
| LU503307A LU503307B1 (en) | 2022-04-18 | 2022-11-02 | Device for automatically preparing ribonucleic acid and preparing method |
| PCT/CN2022/129375 WO2023202033A1 (en) | 2022-04-18 | 2022-11-02 | Automatic rna preparation device and preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210407124.6A CN114703050A (en) | 2022-04-18 | 2022-04-18 | Automatic preparation device and preparation method of RNA |
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| CN114703050A true CN114703050A (en) | 2022-07-05 |
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| CN202210407124.6A Pending CN114703050A (en) | 2022-04-18 | 2022-04-18 | Automatic preparation device and preparation method of RNA |
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| Country | Link |
|---|---|
| CN (1) | CN114703050A (en) |
| LU (1) | LU503307B1 (en) |
| WO (1) | WO2023202033A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023202033A1 (en) * | 2022-04-18 | 2023-10-26 | 湖北文理学院 | Automatic rna preparation device and preparation method |
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Also Published As
| Publication number | Publication date |
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| LU503307B1 (en) | 2024-02-27 |
| LU503307A1 (en) | 2023-10-30 |
| WO2023202033A1 (en) | 2023-10-26 |
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