CN111996113B - Magnetic bead method nucleic acid extraction device and nucleic acid extraction method thereof - Google Patents
Magnetic bead method nucleic acid extraction device and nucleic acid extraction method thereof Download PDFInfo
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- CN111996113B CN111996113B CN202010831922.2A CN202010831922A CN111996113B CN 111996113 B CN111996113 B CN 111996113B CN 202010831922 A CN202010831922 A CN 202010831922A CN 111996113 B CN111996113 B CN 111996113B
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Abstract
The invention discloses a magnetic bead method nucleic acid extraction device and a nucleic acid extraction method thereof. The invention comprises a heating oscillation device, an actuating mechanism frame, a magnetic frame movement mechanism, a sheath movement mechanism and a heating oscillation and refrigeration device; the heating oscillation device and the sheath movement mechanism are combined to crack the reagent on the cracking station; then, the magnetic bead adsorption and magnetism abandoning are realized by combining the executing mechanism frame, the magnetic frame moving mechanism and the sheath moving mechanism; then, the heating oscillation device and the sheath movement mechanism are combined to mix and stir the cracking station, and magnetic beads attached with nucleic acid are adsorbed; then, the executing mechanism frame, the magnetic frame moving mechanism and the sheath moving mechanism are used in combination to wash nucleic acid; and finally, combining the executing mechanism frame, the magnetic frame moving mechanism, the sheath moving mechanism and the heating, vibrating and refrigerating device to elute the nucleic acid. The invention has uniform mixing, reduces the volatilization of aerosol and ensures the length and the integrity of the nucleic acid fragment in the extracted product.
Description
Technical Field
The invention belongs to the technical field of biological nucleic acid extraction and purification, and particularly relates to a magnetic bead method nucleic acid extraction device and a nucleic acid extraction method thereof.
Background
Nucleic acid is a carrier of genetic information, is the most important biological information molecule and is the main object of molecular biology research, therefore, the extraction of nucleic acid is the most important and basic operation in molecular biology experimental technology. With the rapid development of molecular biology technology, the research and analysis of nucleic acid are continuously popularized and applied in the fields of clinical diagnosis, environmental detection, food safety detection and the like, and play a great role.
At present, the automatic nucleic acid extraction instruments on the market mostly adopt a magnetic bar method and a rotation method. Firstly, the magnetic rod method adopts a sheath nested on each magnetic rod, nucleic acid particle magnetic beads absorbed on the surface are absorbed on the lower surface of the sheath through magnetic attraction, the reaction reagents are transferred, and meanwhile, the reagent liquid is rapidly mixed and stirred by the up-and-down high-frequency reciprocating motion of the motor electric magnetic rods and the sheath, so that complex nucleic acid extraction processes such as cracking, adsorption, magnetism abandonment, washing and elution are realized. However, the mixing mode of the sheath moving up and down in a reciprocating manner at high frequency easily generates liquid splashing, the low recovery rate of magnetic beads and the risk of cross contamination of samples are easily caused, and meanwhile, the up and down high frequency reciprocating movement can generate strong vibration on nucleic acid, the length of DNA and RNA can be interrupted, and the integrity of nucleic acid fragments cannot be ensured; secondly, the rotation method adopts a jacket with edges and wings, an independent jacket grabbing mechanism and a transmission mechanism are adopted to enable each jacket to independently rotate at a high speed, so that stirring is realized, the magnetic bead with adsorbed nucleic acid is moved by inserting a magnetic rod into the jacket, and complex nucleic acid extraction processes such as cracking, adsorption, magnetism abandoning, washing, elution and the like are realized by moving the jacket. Therefore, it is imperative to develop an automatic extraction device and method which is more efficient, can reduce the cross-contamination rate of the space, and has simple structure and high stability.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides a magnetic bead method nucleic acid extraction device and a nucleic acid extraction method thereof.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention relates to a magnetic bead method nucleic acid extraction device, which comprises an integral frame, an actuating mechanism, a deep hole plate mounting frame, a heating oscillation device and a heating oscillation and refrigeration device; the integral frame is provided with more than four stations which are arranged side by side, wherein three stations are respectively a cracking station, a magnetic bead station and an elution station, and the rest stations are washing stations; the heating oscillation device is arranged at the cracking station, and the heating oscillation and refrigeration device is arranged at the elution station; deep hole plate mounting frames are fixed at a magnetic bead station and an elution station of the integral frame; a first position sensor is arranged at each station; and the signal output end of the first position sensor is connected with the controller.
The executing mechanism consists of an executing mechanism rack, a driving mechanism I, a magnetic frame moving mechanism, a sheath moving mechanism and an anti-dripping plate moving mechanism; the executing mechanism frame and the slide rail form a sliding pair; the slide rail is parallel to the arrangement direction of each station; the first driving mechanism is controlled by the controller, and drives the executing mechanism rack, the magnetic force frame moving mechanism, the sheath moving mechanism and the anti-dripping plate moving mechanism which are installed on the executing mechanism rack to synchronously move along the arrangement direction of each station. The magnetic frame motion mechanism comprises a driving mechanism II, a magnetic frame upper rack, a magnetic bar and a magnetic frame lower rack; the lower rack of the magnetic frame is provided with n through holes which are arranged in a rectangular shape, wherein n is 96, and each through hole supports one magnetic rod; the upper magnetic frame and the lower magnetic frame are fixed and driven by the second driving mechanism to move up and down; the second driving mechanism is controlled by the controller. The sheath movement mechanism consists of a driving mechanism III, a sheath frame and a sheath; the sheath frame is positioned right below the lower rack of the magnetic frame; the sheath frame is provided with n through holes which are arranged into a rectangle, and the n through holes are aligned with the n magnetic rods one by one; the sheath is provided with n single-hole sheaths which are integrally formed and arranged in a rectangular shape; the sheath is detachably and fixedly connected with the sheath frame, and the n single-hole sheaths are aligned with the n through holes of the sheath frame one by one; the sheath frame is driven by a driving mechanism III to move up and down; the driving mechanism III is controlled by the controller; the anti-dripping plate movement mechanism consists of an anti-dripping plate, a position sensor II and a driving mechanism IV; the driving mechanism IV is controlled by the controller and drives the anti-dripping plate to move along the direction parallel to the sliding rail; the second position sensor is arranged at the initial position of the anti-dripping plate; a third position sensor is arranged at the initial position of the rack on the magnetic frame, and a fourth position sensor is arranged at the initial position of the sheath frame; and signal output ends of the second position sensor, the third position sensor and the fourth position sensor are connected with the controller.
The heating oscillation device and the heating oscillation and refrigeration device respectively comprise a brushless motor, an eccentric block, an oscillation bracket and a joint bearing seat; the base of the brushless motor is fixed on the integral frame; the eccentric block is fixed on an output shaft of the brushless motor; the oscillation bracket is supported on an output shaft of the brushless motor through a rolling bearing and is fixed with the eccentric block; the joint bearing block is supported outside the eccentric block through a joint bearing; the brushless motor is controlled by a controller. The heating oscillation device also comprises a cracking heating aluminum module and a heating film; the pressing plate is fixed with the pressing frame, the heat insulation pad is arranged on the pressing plate, the heating film is arranged on the heat insulation pad, and the cracking heating aluminum module is arranged on the heating film; the pressing frame presses the cracking heating aluminum module and is fixedly connected with the oscillating support through screws; a first temperature sensor is arranged on the cracking heating aluminum module; the signal output end of the first temperature sensor is connected with the controller; the heating film is controlled by a controller. The heating, vibrating and refrigerating device also comprises an eluting and heating aluminum module, a TE refrigerating sheet, a radiator and a fan; the elution heating aluminum module, the TE refrigerating sheet and the radiator are all fixed on a vibration bracket of the heating vibration and refrigerating device; the TE refrigeration sheet is arranged on the radiator, and the elution heating aluminum module is arranged on the TE refrigeration sheet; a temperature sensor II is arranged on the elution heating aluminum module; the signal output end of the temperature sensor II is connected with the controller; an air inlet duct is arranged at the air inlet of the fan, an air duct opening is arranged at the air outlet, and the air duct opening is arranged right opposite to the radiator; the TE refrigeration piece and the fan are controlled by the controller.
Preferably, the N pole and S pole directions of adjacent magnetic bars are different along the length direction or width direction of the rectangle formed by the N through holes.
Preferably, the magnetic rod consists of a permanent magnet round rod and a stainless steel round rod; the permanent magnet round bar is fixedly nested in the hollow end of the stainless steel round bar; the hollow end of the stainless steel round bar is provided with an exhaust pinhole.
Preferably, a buffer pad is arranged between the upper magnetic frame and the lower magnetic frame.
Preferably, the bottom surface of the sheath frame is fixedly provided with three flanges at the outer edge of a rectangle formed by n through holes, wherein two flanges are arranged in parallel with the length direction of the rectangle, and the third flange is arranged in parallel with the width direction of the rectangle; the bottom surface of the sheath frame is provided with a clamping groove at the position of the outer edge of the rectangular width edge without the flange, and the top surface of the sheath frame is provided with two clamping grooves arranged at intervals at the same end position of the clamping groove; a guide groove is formed between each of the two flanges arranged in parallel and the sheath rack, and a limit groove is formed between the other flange and the sheath rack; one end of the sheath is provided with an elastic buckle and an anti-deformation hook which are integrally formed; the deformation-preventing hooks are arranged on two sides of the elastic buckle respectively, and the distance between the two deformation-preventing hooks is equal to the distance between the two buckle grooves. Two side parts of the sheath are respectively embedded into one guide groove at the corresponding side of the sheath frame, and one end of the sheath, which is not provided with the elastic buckle, is embedded into a limit groove of the sheath frame; the elastic buckle of the sheath is buckled with the clamping groove of the sheath frame; the two anti-deformation hooks of the sheath are respectively buckled with the two buckling grooves of the sheath frame.
Preferably, the heating oscillation device and the heating oscillation and refrigeration device further comprise hall sensors for detecting the position of the rotor of the brushless motor; and the signal output end of the Hall sensor is connected with the controller.
Preferably, the deep hole plate mounting rack, the cracking heating aluminum module and the elution heating aluminum module are all provided with n placing holes which are arranged in a rectangular shape, and the placing holes are used for being embedded into the deep hole plate.
The nucleic acid extraction method of the magnetic bead method nucleic acid extraction device comprises the following specific steps:
placing deep hole plates on a cracking heating aluminum module, an eluting heating aluminum module and each deep hole plate mounting rack; injecting reagent and lysate into the deep-hole plate on the cracking heating aluminum module, injecting magnetic bead solution into the deep-hole plate on the deep-hole plate mounting rack at the magnetic bead station, injecting washing buffer solution into the deep-hole plate on the rest deep-hole plate mounting racks, and injecting elution buffer solution into the deep-hole plate on the elution heating aluminum module.
Step two, heating the heating film of the heating oscillation device, after the heating film and the temperature sensor jointly act and feed back and adjust to make the temperature of the cracking heating aluminum module reach the preset temperature, carrying out reagent cracking by adopting one of the following three modes: firstly, a brushless motor of a heating oscillation device drives an eccentric block to enable a cracking heating aluminum module to eccentrically oscillate, a deep hole plate on the cracking heating aluminum module is stopped after eccentric oscillation of the cracking heating aluminum module for a preset time, and a reagent is cracked; secondly, the sheath of the sheath moving mechanism moves downwards into a deep hole plate on the cracking heating aluminum module under the driving of the driving mechanism III, and stops after moving upwards and downwards in the deep hole plate for preset time; then, the sheath moves upwards for resetting; and the sheath moves up and down in the deep hole plate on the cracking heating aluminum module and the cracking heating aluminum module vibrates eccentrically and alternately.
Step three, stopping the motion of the anti-dripping plate when the anti-dripping plate moves to the position right below the sheath under the driving of the driving mechanism four; an executing mechanism rack of the executing mechanism, the magnetic force rack moving mechanism, the sheath moving mechanism and the anti-dripping plate moving mechanism are driven by a driving mechanism I to move synchronously, and after a position sensor I of a magnetic bead station detects that the executing mechanism rack is in place, the executing mechanism rack stops moving, so that a magnetic force rod of the magnetic force rack moving mechanism and a sheath of the sheath moving mechanism stay above a deep hole plate of the magnetic bead station; then, the anti-dripping plate is moved out from the lower part of the sheath, and the anti-dripping plate stops moving after the position sensor II detects that the anti-dripping plate is in place; the magnetic bar of the magnetic frame motion mechanism is inserted into the sheath of the sheath motion mechanism under the drive of the driving mechanism II; then, the magnetic rod of the magnetic rack motion mechanism and the sheath of the sheath motion mechanism synchronously move downwards to the deep hole plate at the magnetic bead station to adsorb the magnetic beads; then, the magnetic rod of the magnetic frame motion mechanism and the sheath of the sheath motion mechanism are lifted and reset synchronously, and the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism; then, the actuating mechanism frame moves under the driving of the driving mechanism I, and after a position sensor I of the cracking station detects that the actuating mechanism frame is in place, the actuating mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate move right above the cracking station; the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the sheath moves downwards and is inserted into a deep hole plate of the cracking station, and magnetism is abandoned; then, the sheath moves upwards to reset; mixing and stirring are carried out by adopting one of the following three ways: firstly, stopping cracking heating aluminum module after eccentric oscillation for preset time; moving the sheath downwards into a deep hole plate on the cracking heating aluminum module, moving the sheath upwards and downwards in the deep hole plate for preset time, and stopping; then, the sheath moves upwards for resetting; and the sheath moves up and down in the deep hole plate on the cracking heating aluminum module and the cracking heating aluminum module vibrates eccentrically and alternately. After the mixing and stirring are finished, the heating film stops heating, and the magnetic rod and the sheath synchronously move downwards to a deep hole plate on the cracking heating aluminum module to adsorb magnetic beads attached with nucleic acid; and finally, the magnetic rod and the sheath are lifted and reset synchronously.
Step four, the sheath is sleeved into the deep hole plate of each washing station in sequence to wash the nucleic acid.
Step five, the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism four; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I of the elution station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate are stopped right above the deep hole plate of the elution station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, and the magnetic rod and the sheath synchronously move downwards into a deep hole plate of an elution station; the TE refrigeration piece of heating oscillation and refrigerating plant gives elution heating aluminium module heating, and after two combined action feedback adjustment of TE refrigeration piece and temperature sensor made the temperature of elution heating aluminium module reach preset temperature, adopt one of following three kinds of modes to elute the nucleic acid to the sheath: firstly, a brushless motor of a heating oscillation and refrigeration device drives an eccentric block to enable an elution heating aluminum module to eccentrically oscillate, and a deep hole plate on the elution heating aluminum module is stopped after eccentric oscillation of the elution heating aluminum module for a preset time; moving the magnetic rod and the sheath up and down in a deep hole plate on the elution heating aluminum module for a preset time, and then stopping; then, the magnetic bar and the sheath move upwards for resetting; and thirdly, the elution heating aluminum module performs eccentric oscillation and moves up and down alternately in a deep hole plate on the elution heating aluminum module through a magnetic rod and a sheath.
Preferably, the process of washing nucleic acid is as follows: the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism IV; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I corresponding to the washing station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate are stopped right above the deep hole plate corresponding to the washing station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the magnetic rod and the sheath synchronously move downwards to the deep hole plate corresponding to the washing station and move up and down in the deep hole plate to wash nucleic acid; after a preset time, the magnetic rod and the sheath are reset upwards synchronously.
Preferably, after nucleic acid elution is finished, the TE refrigerating sheet is controlled to refrigerate the elution heating aluminum module, the fan is started at the same time, the TE refrigerating sheet transmits the heat energy of the elution heating aluminum module to the radiator, and air at an air outlet of the fan is blown to the radiator through an air duct opening to cool the radiator; the TE refrigeration sheet, the fan and the temperature sensor act together to perform feedback adjustment, so that the temperature of the elution heating aluminum module reaches a preset temperature, and the nucleic acid is stored at the preset temperature.
Compared with the prior art, the invention has the beneficial effects that:
1. the lysis, mixing and elution of the present invention can be performed in one of the following ways: 1. stirring up and down independently; 2. independently stirring by high-frequency oscillation; 3. stirring by combining up and down stirring and high-frequency shaking. Therefore, the invention can meet different reagent characteristics, particularly adopts a high-frequency oscillation stirring mode for cracking and elution, is uniformly mixed, reduces the volatilization of aerosol, and can ensure the length and the integrity of the nucleic acid fragment in the extracted product.
2. The anti-dripping plate below the sheath avoids cross contamination when the carrier with nucleic acid and magnetic beads is adsorbed for moving.
3. The invention can realize the full flow of stirring, magnetic attraction, moving, heating, automatic nucleic acid extraction and cold storage, and has high automation degree.
Drawings
FIG. 1-1 is a side view of the whole structure of a nucleic acid extraction apparatus using a magnetic bead method according to the present invention;
FIG. 1-2 is a top view of FIG. 1-1;
FIGS. 1-3 are schematic illustrations of the arrangement of the various stations of the present invention;
FIG. 2-1 is a side view of the actuator of the present invention;
FIG. 2-2 is a left side view of FIG. 2-1;
FIG. 2-3 is a top view of FIG. 2-1;
FIG. 3-1 is an assembled cross-sectional view of the upper frame, the cushion pad, the magnetic rod and the lower frame of the magnetic frame according to the present invention;
FIG. 3-2 is a top view of FIG. 3-1;
3-3 are schematic structural views of the magnetic rod of the present invention;
FIG. 4 is a sectional view of the heating oscillating device according to the present invention;
FIG. 5 is a sectional view of the heating, oscillating and cooling apparatus according to the present invention;
FIG. 6-1 is an assembled perspective view of the sheath holder and sheath of the present invention;
FIG. 6-2 is a top schematic view of the present invention after assembly of the sheath stand and sheath;
fig. 6-3 are end views of the present invention after assembly of the sheath holder and sheath;
FIGS. 6-4 are top views of the sheath stand of the present invention;
FIG. 6-5 is a right side view of FIG. 6-4;
FIGS. 6-6 are schematic end views of the sheath of the present invention after inversion;
in the figure: 1. an actuator; 2. a first position sensor; 3. a deep hole plate mounting rack; 4. a magnetic frame motion mechanism; 5. a sheath movement mechanism; 6. a drip-proof plate movement mechanism; 7. the magnetic frame is arranged on the machine frame; 8. a cushion pad; 9. a magnetic bar; 10. a magnetic frame lower frame; 11. an exhaust pinhole; 12. a cross countersunk head screw; 13. a sheath frame; 14. blocking edges; 15. a sheath; 15-1, elastic buckle; 15-2, anti-deformation hooks; 16. a fastening groove; 17. a clamping groove; 18. a first station; 19. a second station; 20. a third station; 21. a fourth station; 22. a fifth station; 23. a sixth station; 24. cracking and heating the aluminum module; 25. heating the film; 26. a heat insulating pad; 27. oscillating the support; 28. a Hall sensor; 29. an eccentric block; 30. a rolling bearing; 31. a brushless motor; 32. a balancing weight; 33. a knuckle bearing; 34. a joint bearing seat; 35. an air inlet duct; 36. a fan; 37. an air duct opening; 38. eluting and heating the aluminum module; 39. TE refrigeration pieces; 40. a heat sink.
Detailed Description
The technical solution of the embodiment of the present invention will be clearly and completely described below with reference to the accompanying drawings of the present invention.
The magnetic bead method nucleic acid extraction device and the nucleic acid extraction method thereof disclosed by the invention uniformly mix the sample and the extracted reagent by adopting a high-frequency eccentric oscillation mode (certainly, a low-frequency eccentric oscillation mode can also be realized), and simultaneously cooperate with vertical and horizontal motions to uniformly mix, so that the volatilization of aerosol is reduced, and the length and the integrity of a nucleic acid fragment in an extracted product can be ensured.
As shown in fig. 1-1, 1-2, and 1-3, the apparatus for extracting nucleic acid by a magnetic bead method according to an embodiment of the present invention includes an integral rack, an actuator 1, a deep well plate mounting rack 3, a heating oscillation device, and a heating oscillation and refrigeration device; the whole frame is provided with six stations which are arranged side by side, the first station 18 is a cracking station, the second station 19 is a magnetic bead station, the third station 20, the fourth station 21 and the fifth station 22 are washing stations, and the sixth station 23 is an elution station; the heating oscillation device is arranged at the first station, and the heating oscillation and refrigeration device is arranged at the sixth station; the deep hole plate mounting frames 3 are fixed at the second station, the third station, the fourth station and the fifth station of the integral frame; a first position sensor 2 is arranged at each station; and the signal output end of the first position sensor 2 is connected with the controller.
As shown in fig. 2-1, 2-2 and 2-3, the actuator 1 is composed of an actuator frame, a first driving mechanism, a magnetic frame moving mechanism 4, a sheath moving mechanism 5 and an anti-dripping plate moving mechanism 6; the executing mechanism frame and the slide rail form a sliding pair; the slide rail is parallel to the arrangement direction of the six stations; the first driving mechanism is controlled by the controller, and drives the frame of the executing mechanism to synchronously move along the arrangement directions of six stations together with the magnetic frame moving mechanism 4, the sheath moving mechanism 5 and the anti-dripping plate moving mechanism 6 which are arranged on the frame of the executing mechanism; as a preferred embodiment, the first driving mechanism adopts a first driving motor to drive a first synchronous belt transmission mechanism, a synchronous belt of the first synchronous belt transmission mechanism is fixed with a frame of an actuating mechanism, and a driving belt wheel and a driven belt wheel of the first synchronous belt transmission mechanism are both hinged on the whole frame; the first driving motor is controlled by the controller.
As shown in fig. 3-1, 3-2 and 3-3, the magnetic rack movement mechanism 4 comprises a second driving mechanism, an upper magnetic rack frame 7, a buffer cushion 8, a magnetic bar 9 and a lower magnetic rack frame 10; the lower rack 10 of the magnetic frame is provided with n through holes which are arranged in a rectangular shape, wherein n is 96, and each through hole supports one magnetic rod 9; the N pole and S pole directions of the adjacent magnetic rods 9 are different along the length direction or the width direction; the buffer pad 8 is arranged on the lower rack 10 of the magnetic rack; the upper magnetic frame rack 7 is arranged on the buffer pad 8 and is fixed with the lower magnetic frame rack 10; as a preferred embodiment, the upper magnetic frame 7 is connected with the lower magnetic frame 10 through a cross countersunk head screw 12, which is beneficial to replacing and maintaining the magnetic rod 9; the cushion pad 8 plays a role in clearance elimination and prepressing; as a preferred embodiment, the magnetic rod 9 is composed of a permanent magnet round rod and a stainless steel round rod; the permanent magnet round bar is fixedly nested in the hollow end of the stainless steel round bar; the hollow end of the stainless steel round rod is provided with an exhaust pinhole 11, so that the hollow end is prevented from expanding and deforming when the permanent magnet round rod and the stainless steel round rod are pressed and mounted; a third position sensor is arranged at the initial position of the upper rack of the magnetic rack, and the signal output end of the third position sensor is connected with the controller; the upper frame 7 of the magnetic frame is driven by the second driving mechanism to move up and down; the second driving mechanism is controlled by the controller; as a preferred embodiment, the driving mechanism II adopts a driving motor II to drive a synchronous belt transmission mechanism II, and a driving belt wheel and a driven belt wheel of the synchronous belt transmission mechanism II are both hinged on the frame of the actuating mechanism; a driven belt wheel of the synchronous belt transmission mechanism II is fixed with the screw rod I; the first nut block and the first screw form a screw pair, and form a sliding pair with the frame of the actuating mechanism; the upper rack 7 of the magnetic frame is fixed with the first nut block; the second driving motor is controlled by the controller.
As shown in fig. 6-1, 6-2 and 6-3, the sheath moving mechanism 5 is composed of a driving mechanism three, a sheath holder 13 and a sheath 15; as shown in fig. 2-1 and 2-2, the sheath holder 13 is located right below the magnet holder lower frame 10; as shown in fig. 6-2, 6-4 and 6-5, the sheath frame is provided with n through holes arranged in a rectangular shape, and the n through holes are aligned with the n magnetic rods 9 one by one; three flanges 14 are fixedly arranged at the outer edge of a rectangle formed by arranging the n through holes on the bottom surface of the sheath frame, wherein two flanges are arranged in parallel with the length direction of the rectangle, and the third flange is arranged in parallel with the width direction of the rectangle; in a preferred embodiment, the end parts of two flanges parallel to the length direction of the rectangle are respectively contacted with the two ends of a third flange; the bottom surface of the sheath frame is provided with a clamping groove 17 at the position of the outer edge of the rectangular width edge without the flange, and the top surface of the sheath frame is provided with two clamping grooves 16 arranged at intervals at the same end position of the clamping groove 17; a guide groove is formed between each of the two flanges arranged in parallel and the sheath rack, and a limit groove is formed between the other flange and the sheath rack; the guide groove plays a role in guiding and supporting the sheath 15, and can prevent the sheath 15 from deforming; the limiting groove plays a role in limiting and supporting the sheath 15; the locking groove 17 plays a role in fixing and preventing the sheath 15 from moving, and the buckling groove 16 plays a role in fixing the sheath 15 and preventing deformation of the hook. As shown in fig. 6-1 and 6-6, the sheath 15 is provided with n single-hole sheaths which are integrally formed and arranged in a rectangular shape, and an elastic buckle 15-1 and a deformation preventing hook 15-2 which are integrally formed are arranged at one end; the n single-hole sheaths are aligned with the n through holes of the sheath rack one by one; the deformation-preventing hooks are arranged on two sides of the elastic buckle respectively, and the distance between the two deformation-preventing hooks is equal to the distance between the two buckle grooves 16. As shown in fig. 6-1, 6-2 and 6-3, two side parts of the sheath are respectively embedded into a guide groove at the corresponding side of the sheath rack, and one end of the sheath, which is not provided with the elastic buckle 15-1, is embedded into a limit groove of the sheath rack; the elastic buckle 15-1 of the sheath is buckled with the clamping groove 17 of the sheath frame, so that the fixing and limiting effects on the sheath 15 are achieved, and the sheath 15 is prevented from moving; the two anti-deformation hooks 15-2 of the sheath are respectively buckled with the two buckling grooves 16 of the sheath frame, so that two positions of the front end of the sheath 15 are fixed, and the effect of preventing the front end of the sheath 15 from deforming is achieved. A position sensor IV is arranged at the initial position of the sheath frame, and the signal output end of the position sensor IV is connected with the controller; the sheath frame 13 is driven by the driving mechanism III to move up and down; the driving mechanism III is controlled by the controller; as a preferred embodiment, the third driving mechanism adopts a third driving motor to drive a third synchronous belt transmission mechanism, and a driving belt wheel and a driven belt wheel of the third synchronous belt transmission mechanism are both hinged on the frame of the executing mechanism; a driven belt wheel of the synchronous belt transmission mechanism III is fixed with the screw rod II; the nut block II and the screw rod II form a screw pair and form a sliding pair with the executing mechanism frame; the sheath frame 13 is fixed with the nut block II; and the driving motor III is controlled by the controller.
As shown in fig. 2-1, 2-2 and 2-3, the anti-dripping plate movement mechanism 6 is composed of an anti-dripping plate, a position sensor II and a driving mechanism IV; the driving mechanism IV is controlled by the controller and drives the anti-dripping plate to move along the direction parallel to the sliding rail; as a preferred embodiment, the driving mechanism IV drives a synchronous belt transmission mechanism IV by adopting a driving motor IV, and a synchronous belt of the synchronous belt transmission mechanism IV is fixed with the anti-dripping plate; a driving belt wheel and a driven belt wheel of the synchronous belt transmission mechanism IV are both hinged on the frame of the actuating mechanism; the driving motor IV is controlled by the controller; the position sensor II is arranged at the initial position of the anti-dripping plate; the signal output end of the position sensor II is connected with the controller; when the anti-dripping plate moves to the position right below the sheath moving mechanism 5, the pollution among samples can be separated.
As shown in fig. 4 and 5, each of the heating oscillating device and the heating oscillating and cooling device includes a brushless motor 31, an eccentric block 29, an oscillating bracket 27 and a joint bearing block 34; the base of the brushless motor 31 is fixed on the integral frame; the eccentric block 29 is fixed on the output shaft of the brushless motor 31; the oscillating bracket 27 is supported on an output shaft of the brushless motor 31 through a rolling bearing 30 and is fixed with the eccentric block 29; the joint bearing seat 34 is supported outside the eccentric block 29 through a joint bearing 33; the brushless motor 31 is controlled by a controller; the joint bearing seat 34 eccentrically swings to realize oscillation, the joint bearing seat 34 enhances the stability of oscillation, and the reagent sample vortex type uniform oscillation is achieved. As a preferred embodiment, the heating oscillating device and the heating oscillating and refrigerating device further comprise hall sensors 28; the Hall sensor 28 detects the position of the rotor of the brushless motor, the signal output end of the Hall sensor 28 is connected with the controller, and the controller controls the rotor of the brushless motor to be accurately positioned. In a preferred embodiment, a weight 32 is fixed to the joint bearing block 34; the counterweight 32 realizes the integral balance weight of the heating oscillation device and the heating oscillation and refrigeration device.
The heating oscillation device also comprises a cracking heating aluminum module 24 and a heating film 25; the pressing plate is fixed with the pressing frame, the heat insulation pad 26 is arranged on the pressing plate, the heating film 25 is arranged on the heat insulation pad 26, and the cracking heating aluminum module 24 is arranged on the heating film 25; the pressing frame presses the cracking heating aluminum module 24 and is fixedly connected with the oscillating support 27 through screws; a first temperature sensor is arranged on the cracking heating aluminum module 24; the signal output end of the first temperature sensor is connected with the controller; the heating film 25 is controlled by a controller to provide thermal energy to the pyrolysis heating aluminum module 24 to provide the temperature required for pyrolysis. The heating, oscillating and refrigerating device further comprises an eluting and heating aluminum module 38, a TE refrigerating sheet (semiconductor refrigerating sheet) 39, a radiator 40 and a fan 36; the elution heating aluminum module 38, the TE refrigerating sheet 39 and the radiator 40 are all fixed on the oscillating bracket 27 of the heating oscillating and refrigerating device; the TE refrigerating sheet 39 is arranged on the radiator 40, and the elution heating aluminum module 38 is arranged on the TE refrigerating sheet 39; a second temperature sensor is arranged on the eluting and heating aluminum module 38; the signal output end of the temperature sensor II is connected with the controller; an air inlet duct 35 is arranged at an air inlet of the fan 36, an air duct opening 37 is arranged at an air outlet, and the air duct opening 37 is arranged right opposite to the radiator 40; the TE refrigerating sheet 39 and the fan 36 are controlled by a controller; the TE refrigerating sheet 39 provides heat energy for the elution heating aluminum module 38; when the fan 36 is turned on, the TE cooling fins transfer the heat energy of the eluting and heating aluminum module 38 to the radiator 40, and the air from the air duct 37 carries the heat energy of the radiator 40 to cool the radiator 40. It can be seen that the TE cooling fins 39 provide the temperature required for elution heating, and the TE cooling fins 39, the heat sink 40 and the fan 36 together provide low temperature storage, with the storage temperature set at 4 ℃ as a preferred embodiment.
The deep hole plate mounting frame 3, the cracking heating aluminum module 24 and the elution heating aluminum module 38 are all provided with n placing holes arranged in a rectangular shape, and the placing holes are used for embedding the deep hole plate.
The nucleic acid extraction method of the magnetic bead method nucleic acid extraction device comprises the following specific steps:
placing deep-hole plates on the cracking heating aluminum module 24, the eluting heating aluminum module 38 and each deep-hole plate mounting rack 3 (limiting parts are arranged on the cracking heating aluminum module 24, the eluting heating aluminum module 38 and each deep-hole plate mounting rack 3 to limit the deep-hole plates, and the limiting parts can be spring pieces or springs fixed with positioning beads); reagent and lysate are injected into the deep-hole plate on the cracking heating aluminum module 24, magnetic bead liquid is injected into the deep-hole plate on the deep-hole plate mounting rack 3 at the second station, washing buffer is injected into the deep-hole plates on the rest deep-hole plate mounting racks 3, and elution buffer is injected into the deep-hole plates on the elution heating aluminum module 38.
Step two, heating the heating film 25 of the heating oscillation device, after the heating film 25 and the temperature sensor act together to perform feedback regulation to enable the temperature of the cracking heating aluminum module 24 to reach the preset temperature, carrying out reagent cracking by adopting one of the following three modes: firstly, a brushless motor 31 of a heating oscillation device drives an eccentric block 29 to enable a cracking heating aluminum module 24 to eccentrically oscillate, a deep hole plate on the cracking heating aluminum module 24 stops after eccentric oscillation of the cracking heating aluminum module 24 for a preset time, and a reagent is cracked; secondly, the sheath 15 of the sheath moving mechanism 5 is driven by the driving mechanism III to move downwards into a deep hole plate on the cracking heating aluminum module 24 and stop moving up and down in the deep hole plate for preset time; then, the sheath 15 is moved upwards for restoration; and thirdly, the sheath 15 moves up and down in the deep hole plate on the cracking heating aluminum module 24 and the cracking heating aluminum module 24 vibrates eccentrically and alternately.
Step three, stopping the motion of the anti-dripping plate when the anti-dripping plate moves to the position right below the sheath under the driving of the driving mechanism IV (realized by the fact that the driving motor IV in the driving mechanism IV rotates by a set number of revolutions); an executing mechanism rack of the executing mechanism, the magnetic force rack moving mechanism 4, the sheath moving mechanism 5 and the anti-dripping plate moving mechanism 6 synchronously move under the driving of a driving mechanism I, and after a position sensor I2 of a magnetic bead station detects that the executing mechanism rack is in place, the executing mechanism rack stops moving, so that a magnetic rod 9 of the magnetic force rack moving mechanism 4 and a sheath 15 of the sheath moving mechanism 5 stay above a deep hole plate of the magnetic bead station; then, the anti-dripping plate is moved out from the lower part of the sheath, and the anti-dripping plate stops moving after the position sensor II detects that the anti-dripping plate is in place; the magnetic bar 9 of the magnetic frame motion mechanism 4 is inserted into the sheath of the sheath motion mechanism 5 under the drive of the driving mechanism II; then, the magnetic rod 9 of the magnetic rack motion mechanism 4 and the sheath 15 of the sheath motion mechanism 5 synchronously move downwards to a deep hole plate at a magnetic bead station to adsorb magnetic beads; then, the magnetic rod 9 of the magnetic frame motion mechanism 4 and the sheath 15 of the sheath motion mechanism 5 are synchronously lifted and reset, and the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism four; then, the actuating mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I2 of the cracking station detects that the actuating mechanism frame is in place, the actuating mechanism frame stops moving, so that the magnetic rod 9, the sheath 15 and the anti-dripping plate move right above the cracking station; the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the sheath moves downwards and is inserted into a deep hole plate of the cracking station, and magnetism is abandoned; then, the sheath moves upwards to reset; mixing and stirring are carried out by adopting one of the following three ways: firstly, the cracking heating aluminum module 24 stops after eccentric oscillation for a preset time; secondly, the sheath 15 moves downwards into a deep hole plate on the cracking heating aluminum module 24 and stops after moving upwards and downwards in the deep hole plate for preset time; then, the sheath 15 is moved upwards for restoration; and thirdly, the sheath 15 moves up and down in the deep hole plate on the cracking heating aluminum module 24 and the cracking heating aluminum module 24 vibrates eccentrically and alternately. After the mixing and stirring are finished, the heating film 25 stops heating, and the magnetic rod and the sheath synchronously move downwards to a deep hole plate on the cracking heating aluminum module to adsorb magnetic beads attached with nucleic acid; and finally, the magnetic rod and the sheath are lifted and reset synchronously.
Step four, the sheath 15 is sleeved into the deep hole plates of the third station, the fourth station and the fifth station in sequence to wash nucleic acid, and the washing process is as follows: the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism IV; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I2 corresponding to the washing station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod 9, the sheath 15 and the anti-dripping plate are stopped right above the deep hole plate corresponding to the washing station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the magnetic rod 9 and the sheath 15 synchronously move downwards to the deep hole plate corresponding to the washing station and move up and down in the deep hole plate to wash nucleic acid; after a preset time, the magnetic rod 9 and the sheath 15 are reset upwards synchronously.
Step five, the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism four; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I2 at the sixth station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod 9, the sheath 15 and the anti-dripping plate stay right above the deep hole plate at the sixth station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, and the magnetic rod 9 and the sheath 15 synchronously move downwards into a deep hole plate at a sixth station; the TE refrigeration piece of heating oscillation and refrigerating plant gives elution heating aluminium module 38 heating, and after two combined action feedback adjustment of TE refrigeration piece and temperature sensor made elution heating aluminium module 38's temperature reach preset temperature, adopt one of following three kinds of modes to elute the nucleic acid to sheath 15: firstly, a brushless motor 31 of the heating oscillation and refrigeration device drives an eccentric block 29 to enable an elution heating aluminum module 38 to eccentrically oscillate, and a deep hole plate on the elution heating aluminum module 38 stops after eccentric oscillation of the elution heating aluminum module 38 for a preset time; secondly, the magnetic rod 9 and the sheath 15 move up and down in a deep hole plate on the elution heating aluminum module 38 for a preset time and then stop; then, the magnetic bar 9 and the sheath 15 move upwards for restoration; and thirdly, the eccentric oscillation of the elution heating aluminum module 38 and the up-and-down movement of the magnetic rod 9 and the sheath 15 in the deep hole plate are alternately carried out. After nucleic acid elution is finished, the TE refrigerating sheet can be controlled to refrigerate the elution heating aluminum module 38 as required, the fan 36 is started at the same time, the TE refrigerating sheet transmits the heat energy of the elution heating aluminum module 38 to the radiator 40, and air at the air outlet of the fan 36 is blown to the radiator 40 through the air duct opening 37 to cool the radiator 40; and finally, the TE refrigerating plate, the fan 36 and the temperature sensor act together to perform feedback regulation to enable the temperature of the elution heating aluminum module 38 to reach the preset temperature, so that the nucleic acid is stored at the preset temperature.
It should be understood that: any person skilled in the art can modify or easily conceive the technical solutions described in the foregoing embodiments or equivalent substitutes for some technical features within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the embodiments of the present invention, and they should be construed as being included therein. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. Magnetic bead method nucleic acid extraction element, including whole frame, actuating mechanism and deep hole board mounting bracket, its characterized in that: the device also comprises a heating oscillation device and a heating oscillation and refrigeration device; the integral frame is provided with more than four stations which are arranged side by side, wherein three stations are respectively a cracking station, a magnetic bead station and an elution station, and the rest stations are washing stations; the heating oscillation device is arranged at the cracking station, and the heating oscillation and refrigeration device is arranged at the elution station; deep hole plate mounting frames are fixed at a magnetic bead station and an elution station of the integral frame; a first position sensor is arranged at each station; the signal output end of the first position sensor is connected with the controller;
the executing mechanism consists of an executing mechanism rack, a driving mechanism I, a magnetic frame moving mechanism, a sheath moving mechanism and an anti-dripping plate moving mechanism; the executing mechanism frame and the slide rail form a sliding pair; the slide rail is parallel to the arrangement direction of each station; the first driving mechanism is controlled by the controller, and drives the executing mechanism rack to synchronously move along the arrangement direction of each station together with the magnetic force frame moving mechanism, the sheath moving mechanism and the anti-dripping plate moving mechanism which are arranged on the executing mechanism rack; the magnetic frame motion mechanism comprises a driving mechanism II, a magnetic frame upper rack, a magnetic bar and a magnetic frame lower rack; the lower rack of the magnetic frame is provided with n through holes which are arranged in a rectangular shape, wherein n is 96, and each through hole supports one magnetic rod; the upper magnetic frame and the lower magnetic frame are fixed and driven by the second driving mechanism to move up and down; the second driving mechanism is controlled by the controller; the sheath movement mechanism consists of a driving mechanism III, a sheath frame and a sheath; the sheath frame is positioned right below the lower rack of the magnetic frame; the sheath frame is provided with n through holes which are arranged into a rectangle, and the n through holes are aligned with the n magnetic rods one by one; the sheath is provided with n single-hole sheaths which are integrally formed and arranged in a rectangular shape; the sheath is detachably and fixedly connected with the sheath frame, and the n single-hole sheaths are aligned with the n through holes of the sheath frame one by one; the sheath frame is driven by a driving mechanism III to move up and down; the driving mechanism III is controlled by the controller; the anti-dripping plate movement mechanism consists of an anti-dripping plate, a position sensor II and a driving mechanism IV; the driving mechanism IV is controlled by the controller and drives the anti-dripping plate to move along the direction parallel to the sliding rail; the second position sensor is arranged at the initial position of the anti-dripping plate; a third position sensor is arranged at the initial position of the rack on the magnetic frame, and a fourth position sensor is arranged at the initial position of the sheath frame; the signal output ends of the second position sensor, the third position sensor and the fourth position sensor are connected with the controller;
the heating oscillation device and the heating oscillation and refrigeration device respectively comprise a brushless motor, an eccentric block, an oscillation bracket and a joint bearing seat; the base of the brushless motor is fixed on the integral frame; the eccentric block is fixed on an output shaft of the brushless motor; the oscillation bracket is supported on an output shaft of the brushless motor through a rolling bearing and is fixed with the eccentric block; the joint bearing block is supported outside the eccentric block through a joint bearing; the brushless motor is controlled by the controller; the heating oscillation device also comprises a cracking heating aluminum module and a heating film; the pressing plate is fixed with the pressing frame, the heat insulation pad is arranged on the pressing plate, the heating film is arranged on the heat insulation pad, and the cracking heating aluminum module is arranged on the heating film; the pressing frame presses the cracking heating aluminum module and is fixedly connected with the oscillating support through screws; a first temperature sensor is arranged on the cracking heating aluminum module; the signal output end of the first temperature sensor is connected with the controller; the heating film is controlled by a controller; the heating, vibrating and refrigerating device also comprises an eluting and heating aluminum module, a TE refrigerating sheet, a radiator and a fan; the elution heating aluminum module, the TE refrigerating sheet and the radiator are all fixed on a vibration bracket of the heating vibration and refrigerating device; the TE refrigeration sheet is arranged on the radiator, and the elution heating aluminum module is arranged on the TE refrigeration sheet; a temperature sensor II is arranged on the elution heating aluminum module; the signal output end of the temperature sensor II is connected with the controller; an air inlet duct is arranged at the air inlet of the fan, an air duct opening is arranged at the air outlet, and the air duct opening is arranged right opposite to the radiator; the TE refrigeration piece and the fan are controlled by the controller.
2. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: the N pole and the S pole of the adjacent magnetic bars are different along the length direction or the width direction of the rectangle formed by the N through holes.
3. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: the magnetic bar consists of a permanent magnet round bar and a stainless steel round bar; the permanent magnet round bar is fixedly nested in the hollow end of the stainless steel round bar; the hollow end of the stainless steel round bar is provided with an exhaust pinhole.
4. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: a buffer cushion is arranged between the upper magnetic frame and the lower magnetic frame.
5. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: the bottom surface of the sheath frame is fixedly provided with three flanges at the outer edge of a rectangle formed by arranging n through holes, wherein two flanges are arranged in parallel with the length direction of the rectangle, and the third flange is arranged in parallel with the width direction of the rectangle; the bottom surface of the sheath frame is provided with a clamping groove at the position of the outer edge of the rectangular width edge without the flange, and the top surface of the sheath frame is provided with two clamping grooves arranged at intervals at the same end position of the clamping groove; a guide groove is formed between each of the two flanges arranged in parallel and the sheath rack, and a limit groove is formed between the other flange and the sheath rack; one end of the sheath is provided with an elastic buckle and an anti-deformation hook which are integrally formed; the two anti-deformation hooks are respectively arranged on two sides of the elastic buckle, and the distance between the two anti-deformation hooks is equal to the distance between the two buckle grooves; two side parts of the sheath are respectively embedded into one guide groove at the corresponding side of the sheath frame, and one end of the sheath, which is not provided with the elastic buckle, is embedded into a limit groove of the sheath frame; the elastic buckle of the sheath is buckled with the clamping groove of the sheath frame; the two anti-deformation hooks of the sheath are respectively buckled with the two buckling grooves of the sheath frame.
6. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: the heating oscillation device and the heating oscillation and refrigeration device also comprise Hall sensors for detecting the position of the rotor of the brushless motor; and the signal output end of the Hall sensor is connected with the controller.
7. The apparatus for nucleic acid isolation by magnetic bead method according to claim 1, wherein: the deep hole plate mounting rack, the cracking heating aluminum module and the elution heating aluminum module are all provided with n placing holes which are arranged into a rectangle.
8. The method for extracting nucleic acid using a magnetic bead nucleic acid extraction device according to any one of claims 1 to 7, wherein: the method comprises the following specific steps:
placing deep hole plates on a cracking heating aluminum module, an eluting heating aluminum module and each deep hole plate mounting rack; injecting reagents and a lysis solution into a deep-hole plate on the cracking heating aluminum module, injecting a magnetic bead solution into the deep-hole plate on the deep-hole plate mounting rack at the magnetic bead station, injecting a washing buffer solution into the deep-hole plates on the rest deep-hole plate mounting racks, and injecting an elution buffer solution into the deep-hole plates on the elution heating aluminum module;
step two, heating the heating film of the heating oscillation device, after the heating film and the temperature sensor jointly act and feed back and adjust to make the temperature of the cracking heating aluminum module reach the preset temperature, carrying out reagent cracking by adopting one of the following three modes: firstly, a brushless motor of a heating oscillation device drives an eccentric block to enable a cracking heating aluminum module to eccentrically oscillate, a deep hole plate on the cracking heating aluminum module is stopped after eccentric oscillation of the cracking heating aluminum module for a preset time, and a reagent is cracked; secondly, the sheath of the sheath moving mechanism moves downwards into a deep hole plate on the cracking heating aluminum module under the driving of the driving mechanism III, and stops after moving upwards and downwards in the deep hole plate for preset time; then, the sheath moves upwards for resetting; the sheath moves up and down in a deep hole plate on the cracking heating aluminum module and the cracking heating aluminum module is eccentrically vibrated alternately;
step three, stopping the motion of the anti-dripping plate when the anti-dripping plate moves to the position right below the sheath under the driving of the driving mechanism four; an executing mechanism rack of the executing mechanism, the magnetic force rack moving mechanism, the sheath moving mechanism and the anti-dripping plate moving mechanism are driven by a driving mechanism I to move synchronously, and after a position sensor I of a magnetic bead station detects that the executing mechanism rack is in place, the executing mechanism rack stops moving, so that a magnetic force rod of the magnetic force rack moving mechanism and a sheath of the sheath moving mechanism stay above a deep hole plate of the magnetic bead station; then, the anti-dripping plate is moved out from the lower part of the sheath, and the anti-dripping plate stops moving after the position sensor II detects that the anti-dripping plate is in place; the magnetic bar of the magnetic frame motion mechanism is inserted into the sheath of the sheath motion mechanism under the drive of the driving mechanism II; then, the magnetic rod of the magnetic rack motion mechanism and the sheath of the sheath motion mechanism synchronously move downwards to the deep hole plate at the magnetic bead station to adsorb the magnetic beads; then, the magnetic rod of the magnetic frame motion mechanism and the sheath of the sheath motion mechanism are lifted and reset synchronously, and the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism; then, the actuating mechanism frame moves under the driving of the driving mechanism I, and after a position sensor I of the cracking station detects that the actuating mechanism frame is in place, the actuating mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate move right above the cracking station; the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the sheath moves downwards and is inserted into a deep hole plate of the cracking station, and magnetism is abandoned; then, the sheath moves upwards to reset; mixing and stirring are carried out by adopting one of the following three ways: firstly, stopping cracking heating aluminum module after eccentric oscillation for preset time; moving the sheath downwards into a deep hole plate on the cracking heating aluminum module, moving the sheath upwards and downwards in the deep hole plate for preset time, and stopping; then, the sheath moves upwards for resetting; the sheath moves up and down in a deep hole plate on the cracking heating aluminum module and the cracking heating aluminum module is eccentrically vibrated alternately; after the mixing and stirring are finished, the heating film stops heating, and the magnetic rod and the sheath synchronously move downwards to a deep hole plate on the cracking heating aluminum module to adsorb magnetic beads attached with nucleic acid; finally, the magnetic bar and the sheath are lifted and reset synchronously;
step four, the sheath is sleeved in the deep hole plate of each washing station in sequence to wash nucleic acid;
step five, the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism four; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I of the elution station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate are stopped right above the deep hole plate of the elution station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, and the magnetic rod and the sheath synchronously move downwards into a deep hole plate of an elution station; the TE refrigeration piece of heating oscillation and refrigerating plant gives elution heating aluminium module heating, and after two combined action feedback adjustment of TE refrigeration piece and temperature sensor made the temperature of elution heating aluminium module reach preset temperature, adopt one of following three kinds of modes to elute the nucleic acid to the sheath: firstly, a brushless motor of a heating oscillation and refrigeration device drives an eccentric block to enable an elution heating aluminum module to eccentrically oscillate, and a deep hole plate on the elution heating aluminum module is stopped after eccentric oscillation of the elution heating aluminum module for a preset time; moving the magnetic rod and the sheath up and down in a deep hole plate on the elution heating aluminum module for a preset time, and then stopping; then, the magnetic bar and the sheath move upwards for resetting; and thirdly, the elution heating aluminum module performs eccentric oscillation and moves up and down alternately in a deep hole plate on the elution heating aluminum module through a magnetic rod and a sheath.
9. The method for extracting nucleic acid using a magnetic bead method nucleic acid extraction device according to claim 8, comprising: the process for washing nucleic acid is specifically as follows: the anti-dripping plate stops moving when moving to the position right below the sheath under the driving of the driving mechanism IV; then, the executing mechanism frame moves under the driving of the driving mechanism I, and after the position sensor I corresponding to the washing station detects that the executing mechanism frame is in place, the executing mechanism frame stops moving, so that the magnetic rod, the sheath and the anti-dripping plate are stopped right above the deep hole plate corresponding to the washing station; then, the anti-dripping plate is moved out from the lower part of the sheath, after the position sensor II detects that the anti-dripping plate is in place, the anti-dripping plate stops moving, the magnetic rod and the sheath synchronously move downwards to the deep hole plate corresponding to the washing station and move up and down in the deep hole plate to wash nucleic acid; after a preset time, the magnetic rod and the sheath are reset upwards synchronously.
10. The method for extracting nucleic acid using a magnetic bead method nucleic acid extraction device according to claim 8, comprising: after nucleic acid elution is finished, the TE refrigerating sheet is controlled to refrigerate the elution heating aluminum module, the fan is started at the same time, the TE refrigerating sheet transmits the heat energy of the elution heating aluminum module to the radiator, and air at the air outlet of the fan is blown to the radiator through the air duct opening to cool the radiator; the TE refrigeration sheet, the fan and the temperature sensor act together to perform feedback adjustment, so that the temperature of the elution heating aluminum module reaches a preset temperature, and the nucleic acid is stored at the preset temperature.
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| WO2019000708A1 (en) * | 2017-06-28 | 2019-01-03 | 苏州长光华医生物医学工程有限公司 | Blood type analyzer |
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| CN103897987B (en) * | 2014-02-18 | 2016-05-04 | 中国农业大学 | Nucleic acid automatic extracting device and method thereof based on nanometer magnetic bead |
| CN107058062A (en) * | 2017-06-09 | 2017-08-18 | 苏州天隆生物科技有限公司 | A kind of rotary nucleic acid-extracting apparatus and its control method |
| WO2019000708A1 (en) * | 2017-06-28 | 2019-01-03 | 苏州长光华医生物医学工程有限公司 | Blood type analyzer |
| CN109055364A (en) * | 2018-09-27 | 2018-12-21 | 无锡百泰克生物技术有限公司 | A kind of instrument for extracting nucleic acid and its application method |
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