CN111394220B - Nucleic acid extraction device - Google Patents

Nucleic acid extraction device Download PDF

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Publication number
CN111394220B
CN111394220B CN202010212449.XA CN202010212449A CN111394220B CN 111394220 B CN111394220 B CN 111394220B CN 202010212449 A CN202010212449 A CN 202010212449A CN 111394220 B CN111394220 B CN 111394220B
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test tube
centrifugal
module
nucleic acid
mixing
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CN111394220A (en
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邱宪波
吴杰
吉尚志
叶祥忠
邱子欣
龚士淞
李益民
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Beijing Controls & Standards Biotechnology Co ltd
Beijing University of Chemical Technology
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Beijing Controls & Standards Biotechnology Co ltd
Beijing University of Chemical Technology
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA

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Abstract

The invention discloses a nucleic acid extraction device. The nucleic acid extraction device comprises a base, a centrifugal module and a mixing module, wherein the centrifugal module comprises a centrifugal tray, the centrifugal tray is rotatably arranged on the base around a centrifugal axis, the mixing module comprises a test tube fixing piece, the test tube fixing piece is provided with a test tube accommodating hole for accommodating a test tube, the test tube fixing piece is rotatably arranged on the centrifugal tray around a mixing axis, and the mixing axis is obliquely arranged relative to the centrifugal axis. According to the nucleic acid extraction device, the test tube fixing part rotates relative to the centrifugal tray, and the mixing axis is inclined relative to the centrifugal axis, so that the rotating speed of the test tube fixing part has a component in the vertical direction, and reagents in the test tube can be enabled to perform upside down movement so as to better mix the reagents.

Description

Nucleic acid extraction device
Technical Field
The invention relates to the field of bioengineering, in particular to a nucleic acid extraction device.
Background
Nucleic acids are the most basic substance of life, which is composed of biological macromolecular compounds, and are vital to the whole life system. Nucleic acids have extremely important applications, both in practical use and in scientific research. Many genetic disorders are closely related to nucleic acids. In general, the investigation of nucleic acids is of great significance.
As a basic approach to biology, nucleic acid extraction is the starting point for downstream nucleic acid detection, research analysis, and product development, and is also a key step in nucleic acid diagnostics. Nucleic acid extraction has high requirements on the quality and integrity of the isolated nucleic acids, both for research and for diagnosis, so that rapid, efficient and accurate nucleic acid extraction techniques are a prerequisite for subsequent research. Aqueous two-phase systems are mutually insoluble two-phase or multiphase systems formed by mixing certain polymers or polymers and salts or other combinations in water at certain concentrations. The aqueous two-phase nucleic acid extraction method is a novel nucleic acid extraction technology, and uses the difference of partition coefficients of solutes in two phases to extract target nucleic acid so as to realize the extraction of nucleic acid.
According to the operation requirement of extracting nucleic acid by a double-aqueous phase nucleic acid extraction system, various instruments such as a centrifuge, a heating plate, an oscillator and the like are often needed, and complicated manual operation is carried out. Currently, most nucleic acid extraction instruments employ conventional nucleic acid extraction methods. The whole extraction reaction time is long, the efficiency is low, the operation is complex, the cost is high, and the degree of automation is low.
Disclosure of Invention
The invention aims to provide a nucleic acid extraction device which can mix reagents well.
The present invention provides a nucleic acid extraction device comprising:
a base;
the centrifugal module comprises a centrifugal tray, and the centrifugal tray is rotatably arranged on the base around a centrifugal axis; and
the mixing module comprises a test tube fixing piece, wherein the test tube fixing piece is provided with a test tube accommodating hole for accommodating a test tube, and the test tube fixing piece is rotatably arranged on the centrifugal tray around a mixing axis, and the mixing axis is obliquely arranged relative to the centrifugal axis.
In some embodiments, the mixing axis is inclined at an angle in the range of 15 ° to 45 ° relative to the axis of centrifugation.
In some embodiments, the axis of the tube-receiving bore intersects the mixing axis.
In some embodiments, the axis of the tube-receiving aperture is perpendicular to the mixing axis.
In some embodiments, the cuvette holder is configured to periodically forward and reverse relative to the centrifuge tray.
In some embodiments, the nucleic acid extraction device comprises two mixing modules symmetrically disposed with respect to the axis of centrifugation.
In some embodiments, the centrifugal tray includes a central tray body and inclined tray bodies disposed on two sides of the central tray body, and the two mixing modules are respectively disposed on the two inclined tray bodies correspondingly.
In some embodiments, the mixing module further comprises a housing arranged outside the test tube fixing part, the housing comprises a groove body and a cover body rotationally connected to the groove body, a test tube fastening strip is arranged on the inner wall of the cover body, and when the cover body rotates to a position for closing the groove body, the test tube fastening strip is abutted with a test tube cover of the test tube to compress the test tube cover.
In some embodiments, the test tube fixing member has a plurality of test tube accommodation holes that the interval set up, and the test tube fastening strip has spacing recess, and spacing recess extends in order to make test tube lid card locate in the spacing recess along the direction of distribution of a plurality of test tube accommodation holes.
In some embodiments, an elastomeric material is mounted within the limit groove.
In some embodiments, the test tube holder includes a connecting edge on both sides of the bottom, and the mixing module further includes a pad disposed between the connecting edge and the bottom surface of the housing, the connecting edge being connected to the housing by the pad.
In some embodiments, the nucleic acid extraction apparatus further comprises a heating module comprising a heating element that heats the tube holder.
In some embodiments, the heating element is an electrothermal film, and the electrothermal film is attached to the end surface of the test tube fixing element.
In some embodiments, the heating module further comprises a temperature sensor, and the tube holder further comprises a detection hole disposed between two adjacent tube receiving holes, the temperature sensor being disposed in the detection hole.
In some embodiments, the nucleic acid extraction apparatus further comprises a heat-dissipating module, wherein the heat-dissipating module comprises a side heat-dissipating fan disposed on a side of the cuvette holder; and/or, the heat dissipation module further comprises a bottom surface heat dissipation fan arranged on the bottom surface of the test tube fixing piece.
In some embodiments, the hybrid module includes a housing with a fan heat dissipation aperture disposed therein.
In some embodiments, the nucleic acid extraction apparatus further comprises a control module comprising a primary controller coupled to the centrifugation module, a secondary controller coupled to the mixing module, and an electrically conductive slip ring connected to the centrifugation tray and electrically connecting the primary controller and the secondary controller.
Based on the technical scheme provided by the invention, the nucleic acid extraction device comprises a base, a centrifugal module and a mixing module, wherein the centrifugal module comprises a centrifugal tray, the centrifugal tray is rotatably arranged on the base around a centrifugal axis, the mixing module comprises a test tube fixing piece, the test tube fixing piece is provided with a test tube accommodating hole for accommodating a test tube, the test tube fixing piece is rotatably arranged on the centrifugal tray around a mixing axis, and the mixing axis is obliquely arranged relative to the centrifugal axis. According to the nucleic acid extraction device, the test tube fixing part rotates relative to the centrifugal tray, and the mixing axis is inclined relative to the centrifugal axis, so that the rotating speed of the test tube fixing part has a component in the vertical direction, and reagents in the test tube can be enabled to perform upside down movement so as to better mix the reagents.
Other features of the present invention and its advantages will become apparent from the following detailed description of exemplary embodiments of the invention, which proceeds with reference to the accompanying drawings.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiments of the invention and together with the description serve to explain the invention and do not constitute a limitation on the invention. In the drawings:
FIG. 1 is a schematic diagram showing the structure of a nucleic acid isolation apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of the centrifugal module of FIG. 1;
FIG. 3 is a schematic view of the mixing module and centrifuge tray of FIG. 1;
FIG. 4 is a schematic diagram of a connection structure between the mixing module and the centrifuge tray of FIG. 3;
FIG. 5 is a schematic view of the mixing module and heating module inside the housing of FIG. 3;
FIG. 6 is a schematic view of the heating module of FIG. 5 with the heat dissipating fan removed;
FIG. 7 is a schematic diagram of the control module of FIG. 1;
fig. 8 is a schematic diagram of a control principle of the control module shown in fig. 7.
Each reference numeral represents:
1. a base;
2. a centrifugal module;
21. a centrifugal tray; 211. a central tray body; 212. a tilting tray body; 213. a vertical tray; 22. a DC motor; 221. an output shaft of the DC motor; 23. a coupling; 25. an extension shaft; 26. a shaft sleeve; 27. a motor tray; 28. a motor bracket;
3. a mixing module;
31. a housing; 311. a tank body; 311A, fan heat dissipation holes; 311B, ventilation and heat dissipation holes; 312. a cover body; 32. a test tube fixing member; 321. a test tube accommodation hole; 322. connecting edges; 33. a cushion block; 34. locking; 35. a stepping motor; 36. bearing fixing seat, 37, bearing; 38. a stepping motor shaft sleeve; 39. a test tube fastening strip;
4. a control module;
41. a main controller; 42. a sub-controller; 43. a conductive slip ring;
5. a heating module;
6. a heat dissipation module;
61. a side cooling fan; 62. a bottom surface heat radiation fan.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. The following description of at least one exemplary embodiment is merely exemplary in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The relative arrangement of the components and steps, numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present invention unless it is specifically stated otherwise. Meanwhile, it should be understood that the sizes of the respective parts shown in the drawings are not drawn in actual scale for convenience of description. Techniques, methods, and apparatus known to one of ordinary skill in the relevant art may not be discussed in detail, but should be considered part of the authorization specification where appropriate. In all examples shown and discussed herein, any specific values should be construed as merely illustrative, and not a limitation. Thus, other examples of the exemplary embodiments may have different values. It should be noted that: like reference numerals and letters denote like items in the following figures, and thus once an item is defined in one figure, no further discussion thereof is necessary in subsequent figures.
Spatially relative terms, such as "above … …," "above … …," "upper surface at … …," "above," and the like, may be used herein for ease of description to describe one device or feature's spatial location relative to another device or feature as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations in use or operation in addition to the orientation depicted in the figures. For example, if the device in the figures is turned over, elements described as "above" or "over" other devices or structures would then be oriented "below" or "beneath" the other devices or structures. Thus, the exemplary term "above … …" may include both orientations of "above … …" and "below … …". The device may also be positioned in other different ways and the spatially relative descriptions used herein are construed accordingly.
As shown in fig. 1 to 6, a nucleic acid extraction apparatus according to an embodiment of the present invention includes:
a base 1;
the centrifugal module 2 comprises a centrifugal tray 21, and the centrifugal tray 21 is rotatably arranged on the base 1 around a centrifugal axis; and
the mixing module 3 comprises a test tube holder 32, the test tube holder 32 has a test tube receiving hole 321 for receiving the test tube a, and the test tube holder 32 is rotatably arranged on the centrifugal tray 21 about a mixing axis which is arranged obliquely with respect to the centrifugal axis.
The tube holder 32 of the nucleic acid extraction apparatus according to the embodiment of the present invention rotates with respect to the centrifugal tray 21 and the mixing axis is inclined with respect to the centrifugal axis, so that the rotation speed of the tube holder 32 has a component in the vertical direction to cause the reagent in the tube a to perform an upside down movement to mix the reagent better.
In order to provide a better centrifuging effect for the reagent, the mixing axis of the present embodiment has an inclination angle in the range of 15 ° to 45 ° with respect to the centrifuging axis.
As shown in fig. 3, the mixing module 3 of the present embodiment further includes a housing 31 disposed outside the test tube holder 32 and fixedly connected to the test tube holder 32. The housing 31 has a square structure and includes a groove body 311 and a cover body 312 rotatably connected to the groove body 311. The cover 312 is connected with the slot 311 through the latch 34 to lock the housing 31. Specifically, the lock catch 34 includes a box catch disposed on the slot 311 and a hook disposed on the cover 312, and the hook is engaged with the box catch.
In the following description, the side of the cover 312 is referred to as an upper side, and the side opposite to the cover 312 is referred to as a lower side. As shown in fig. 6, the test tube holder 32 of the present embodiment includes a connection side 322 at both sides of the bottom, and the mixing module 3 further includes a spacer 33 disposed between the connection side 322 and the bottom surface of the housing 31, and the connection side 322 is connected to the housing 31 through the spacer 33.
The mixing module 3 of the present embodiment further includes a stepping motor 35 for driving the housing 31 to rotate. The output shaft of the stepper motor 35 is connected to a stepper motor sleeve 38, and the stepper motor sleeve 38 is connected to the side wall of the housing 31. The stepper motor sleeve 38 is fitted into the inner diameter of the bearing 37 to form a tight fit with the inner ring structure of the bearing 37, the outer ring of the bearing 37 is fitted into the bearing holder 36, and the bearing holder 36 is screwed onto the centrifugal tray 21. The output shaft of the stepping motor drives the mixing module 3 to rotate, so that the function of mixing reagents is achieved. And after the mixing module 3 is fixed, the weight of the mixing module is concentrated on the bearing fixing seat 36, and the stepping motor 35 only plays a role in transmission and cannot bear the weight of the whole mixing module 3.
In order to better mix the reagents, the cuvette holder 32 of the present embodiment is configured to perform periodic forward and reverse rotation with respect to the centrifuge tray 21. Specifically, the output shaft of the stepping motor 35 is controlled to perform periodic forward and reverse rotation, so that the housing 3 and the cuvette holder 32 are driven to perform periodic forward and reverse rotation. As shown in fig. 6, the cuvette holder 32 of the present embodiment has a plurality of cuvette receiving holes 321 arranged at intervals, and the plurality of cuvette receiving holes 321 are sequentially arranged along a straight line, thereby realizing an operation of extracting nucleic acids from a plurality of reagents to improve efficiency.
To further mix the reagents, the axis of the test tube receiving hole 321 of the present embodiment intersects the mixing axis. So set up, thereby reagent in test tube A moves in test tube A's axis direction so that reagent intensive mixing. The cuvette holder 32 of the present embodiment comprises a plurality of cuvette receiving holes 321, wherein the axes of the cuvette receiving holes may be coplanar and intersect with the mixing axis, and the axes of the cuvette receiving holes may not be coplanar and intersect with the mixing axis.
Specifically, the axis of the cuvette receiving hole 321 is perpendicular to the mixing axis. When the axis of the test tube accommodating hole 321 is perpendicular to the mixing axis, the test tube a follows the test tube fixing member 32 to rotate around the mixing axis, and the linear velocity direction of the test tube a is the axis of the test tube accommodating hole 321, so that the reagent velocity in the test tube a is the same, and in the process that the test tube fixing member 32 follows the housing 31 to perform periodic forward and reverse rotation, the reagent in the test tube a is inverted along the axis direction of the test tube a. The arrangement ensures that the reagent has sufficient space movement in the process of mixing upside down, and simultaneously ensures that the reagent is centrifuged at the angle, so that the reagent can obtain better centrifugal effect.
The nucleic acid isolation apparatus of the present embodiment further includes a heating module 5. The heating module 5 includes a heating member that heats the test tube holder 32.
Specifically, the test tube holder 32 of the present embodiment is a metal block. The heating element is an electrothermal film which is attached to the end face of the test tube fixing element 32. Preferably, electrothermal films are attached to both side end surfaces of the tube fixing member 32 of the present embodiment so that the tube accommodating hole 321 is located between the two electrothermal films to ensure the stability of heating. And the electrothermal film covers the surface of the metal block in a large area to improve heating efficiency.
In other embodiments, the heating element may be an electrothermal tube, an electrothermal ceramic plate, or other workpieces that can perform heating function, and is not limited to the manner of heating by using electric energy.
The end face of the test tube fixing member 32 of this embodiment is provided with a mounting groove, and the electrothermal film is embedded into the mounting groove to achieve a good heating effect.
The diameter of the test tube accommodating hole 321 of this embodiment is slightly larger than that of the test tube so that the test tube can have better fit between the test tube wall and the test tube accommodating hole wall to ensure the direct conduction of heat when the test tube is inserted into the test tube accommodating hole 321. Meanwhile, the wall thicknesses between the adjacent test tube accommodating holes are the same so as to ensure the uniformity and stability of heat conduction in the heating process and reduce the temperature difference between the holes.
In order to monitor the heating temperature, the heating module 5 of this embodiment further includes a temperature sensor, and the test tube holder 32 further includes a detection hole disposed between two adjacent test tube accommodating holes, and the temperature sensor is disposed in the detection hole. For example, the detection hole is opened in the middle of the first test tube accommodating hole and the second test tube accommodating hole, and the temperature of the first test tube accommodating hole and the second test tube accommodating hole can be better reflected. Because the electrothermal film is symmetrically arranged and the test tube accommodating holes are also arranged in the middle of the test tube fixing piece, the heating process is symmetrically performed and the temperature transmission is symmetrical, so that the temperature between the first test tube accommodating hole and the second test tube accommodating hole can reflect the temperature between the third test tube accommodating hole and the fourth test tube accommodating hole.
In order to avoid the problem that the reagent leaks due to the fact that the tube cover is pushed open because of the increase of the temperature and the pressure in the tube a during the heating process, the mixing module 3 of the present embodiment further includes a tube fastening strip 39 disposed on the inner wall of the cover 312, and when the cover 312 rotates to the position of the closed groove 311, the tube fastening strip 39 abuts against the tube cover of the tube a to press the tube cover, thereby preventing the tube cover from being detached.
Specifically, the test tube fastening strip 39 of the present embodiment has a limiting groove, and the limiting groove extends along the distribution direction of the plurality of test tube accommodating holes 321 so that the plurality of test tube caps are clamped in the limiting groove.
Elastic materials are arranged in the limiting grooves of the embodiment. When the cover 312 is closed, the elastic material is in contact with the test tube cover, and the elastic material is in contact with the test tube cover and deforms to a certain extent by controlling the thickness of the elastic material so as to continuously apply pressure to the test tube cover to fasten the test tube.
After mixing the reagents, the temperature of the reagents inside the test tube needs to be reduced. In this embodiment, the nucleic acid isolation apparatus further includes a heat dissipation module 6, and the heat dissipation module 6 includes a side heat dissipation fan 61 provided on the side of the heating module 5; and/or the heat dissipation module 6 includes a bottom surface heat dissipation fan 62 provided at the bottom surface of the heating module 5.
As shown in fig. 3, the housing 31 of the present embodiment is provided with a fan heat dissipation hole 311A on a surface corresponding to the heat dissipation fan, specifically, both the side surface and the bottom surface of the housing 31 are provided with the fan heat dissipation hole 311A.
The groove body 311 of the present embodiment is a square structure, and includes four side surfaces and a bottom surface that are circumferentially arranged, wherein one side surface is a connection side surface connected with the centrifugal tray 21, and two opposite side surfaces and the bottom surface are both provided with fan heat dissipation holes 311A. As shown in fig. 3, the lower side of the fan heat dissipation hole 311A is further provided with a bar hole for penetrating a power supply line of the heat dissipation fan and the like to facilitate the arrangement and layout of the line. And the strip-shaped holes can further improve the heat dissipation effect.
In order to further improve the heat dissipation effect, the side of the slot body 311 opposite to the connection side of the present embodiment is further provided with a plurality of ventilation and heat dissipation holes 311B.
In order to maintain the balance of centrifugation, as shown in FIG. 1, the nucleic acid extraction apparatus of the present embodiment includes two mixing modules 3 symmetrically arranged with respect to the axis of centrifugation.
As shown in fig. 2, the centrifugal tray 21 includes a central tray body 211 and inclined tray bodies 212 disposed on both sides of the central tray body 211, and two mixing modules 3 are respectively disposed on the two inclined tray bodies 212 correspondingly.
The centrifugal module 2 of the present embodiment further comprises centrifugal driving means for driving the centrifugal tray 21 to rotate. Specifically, the centrifugal driving device of the present embodiment is a dc motor 22, and an output shaft 221 of the dc motor 22 is connected to an extension shaft 25 through a coupling 23. The end of the extension shaft 25 is sleeved with a shaft sleeve 26, and the shaft sleeve 26 is connected with the centrifugal tray 21.
In this embodiment, as shown in FIGS. 7 and 8, the nucleic acid extraction apparatus further includes a control module 4, the control module 4 including a main controller 41 coupled to the centrifugation module 2, a sub-controller 42 coupled to the mixing module 3, and an electrically conductive slip ring 43, the electrically conductive slip ring 43 being connected to the centrifugation tray 21 and electrically connecting the main controller 41 and the sub-controller 42.
Wherein the main controller 41 is disposed on the base 1.
Since the mixing module 3 and the heating module 5 both rotate along with the centrifugal tray 21, and the sub-controller 42 is used for controlling the mixing module 3 and the heating module 5, in order to reduce the connecting wires between the control module and the mixing module 3 and the heating module 5 and simplify the device, the sub-controller 42 in this embodiment synchronously rotates along with the centrifugal tray 21.
Specifically, the centrifugal tray 21 of the present embodiment further includes a vertical tray body 213 connected to the inclined tray body 212, and the sub controller 42 is connected to the vertical tray body 213 and rotates with the vertical tray body 213.
The structure and operation of the nucleic acid isolation apparatus according to the embodiment of the present invention will be described in detail with reference to FIGS. 1 to 8.
As shown in fig. 1 to 7, the nucleic acid extraction apparatus of the present embodiment includes a base 1, a centrifugation module 2, a mixing module 3, a control module 4, a heating module 5, and a heat dissipation module 6. The base 1 is fixedly arranged, the centrifugal module 2 is rotatably arranged on the base 1, and the mixing module 3 is rotatably arranged on the centrifugal module 2.
The centrifugal module 2 comprises a centrifugal tray 21, a direct current motor 22, a coupling 23, an extension shaft 25, a shaft sleeve 26, a motor tray 27 and a motor bracket 28. Two motor brackets 28 are fixed on the base 1 side by side, and motor tray 27 is connected on the top of motor bracket 8, and the top of direct current motor 22 is connected with motor tray 27 and has certain interval between the bottom of direct current motor 22 and base 1. An output shaft 221 of the dc motor 22 passes through the motor tray 27 and is connected to the extension shaft 25 by a coupling 23. The extension shaft 25 is inserted into a bore of the sleeve 26 and is connected to the sleeve 26 by a threaded connection. The centrifugal tray 21 is fixed to the upper surface of the boss 25. The direct current motor 22 has a wider rotation speed adjusting range, provides larger torque to drive the centrifugal tray 21 to rotate at a low rotation speed, and ensures good centrifugal effect at a high rotation speed.
The nucleic acid extraction apparatus of the present embodiment includes two mixing modules 3 symmetrically arranged with respect to the centrifugal axis so that the centrifugal turntable 21 of the present embodiment is strictly symmetrical left and right, ensuring the stability of centrifugation.
As shown in fig. 3 to 6, the mixing module 3 of the present embodiment includes a housing 31, a test tube holder 32, a pad 33, a lock catch 34, a stepping motor 35, a bearing holder 36, a bearing 37, a stepping motor shaft sleeve 38, and a test tube fastening strip 39.
As shown in fig. 3, the housing 31 includes a groove 311 and a cover 312. The test tube holder 32 is fixedly disposed in the housing 31. As shown in fig. 5, the cuvette holder 32 is provided with a plurality of cuvette receiving holes 321 arranged in order along a straight line, and a cuvette a containing a reagent is placed in the cuvette receiving holes 321. And the test tube holder 32 includes a connecting edge 322 at both sides of the bottom, the connecting edge 322 is connected with the bottom surface of the housing 31, and a spacer 33 is disposed between the connecting edge 322 and the bottom surface of the housing 31, so that a certain distance is provided between the test tube holder 32 and the bottom surface of the housing 31.
As shown in fig. 3, the end surface of the stepping motor 35 is fixed to the lower surface of the centrifugal tray 21 by screws. The output shaft of the stepper motor 35 is placed into the circular hole of the stepper motor sleeve 38. The stepper motor sleeve 38 is fitted into the inner diameter of the bearing 37 and forms a tight fit with the inner ring structure of the bearing 37, the outer ring of the bearing 37 is fitted into the bearing holder 36, and the bearing holder 36 is screwed onto the centrifugal tray 21. The mixing module 3 is driven by a stepper motor 35 to effect circular movement relative to the centrifuge tray 21 via bearings 37 to effect mixing of the reagents. And after the mixing module 3 is fixed, the weight of the mixing module is concentrated on the bearing fixing seat 36, and the stepping motor 35 only plays a role in transmission and cannot bear the weight of the whole mixing module 3.
The housing 31 is driven by the stepping motor 35 to rotate relative to the centrifugal tray 21 to mix the reagents in the test tubes a upside down, and in order to avoid the reagents from spilling out during the mixing process, the mixing module 3 of the present embodiment further includes a test tube fastening strip 39 disposed inside the cover 312, and when the cover 312 rotates relative to the slot 311 to close the slot 311, the test tube fastening strip 3939 presses the tops of the test tubes a to prevent the test tube cover from being detached.
Specifically, the test tube fastening strip 39 of this embodiment is a metal strip with a rectangular groove formed thereon. Rubber strips with certain elasticity are arranged in the grooves. When the cover 312 is closed, the test tube fastening strip 39 comes into contact with the test tube fixed in the test tube holder 32. In the actual test process, the thickness of the rubber strip is controlled to enable the rubber strip to be in contact with the test tube cover to generate certain deformation, and the test tube cover is continuously pressurized to fasten the test tube. Meanwhile, the risk that the reagent leaks when the test tube cover is jacked up by the steam generated by the gas heated at high temperature in the test tube is reduced.
The nucleic acid isolation apparatus of this embodiment further includes a heating module 5, and the heating module 5 includes a heating member connected to the test tube holder 32, and the heating member heats the test tube A by the test tube holder 32.
Specifically, the heating element in this embodiment is an electrothermal film, and electrothermal films are adhered to two sides of the test tube fixing element 32. The test tube fixing member 32 of this embodiment is of a block structure, and electric heating films are respectively embedded on two end surfaces of the test tube fixing member, and the electric heating films are arranged front and back, so that the surface of the test tube fixing member 32 is covered in a large area, and the test tube accommodating hole 321 is clamped between the two electric heating films, thereby ensuring the heating stability.
The test tube fixing piece 32 of this embodiment is further provided with a detection hole, a temperature sensor is placed in the detection hole, and the inside of the detection hole is filled with heat-conducting silicone grease, so that the temperature in the detection hole can be guaranteed to follow the temperature of the test tube fixing piece 32 better. The detection hole is opened in the middle of two adjacent test tube accommodation holes, can better reflect the temperature of two test tube accommodation holes.
The heat radiation module 6 of the present embodiment includes a side heat radiation fan 61 provided on the side of the cuvette holder 32 and a bottom heat radiation fan 62 provided on the bottom of the cuvette holder 32. The housing 31 of the present embodiment is provided with a fan heat radiation hole 311A at a position corresponding to the heat radiation fan. The side cooling fan 61 blows air outward from the housing 31. The bottom surface heat radiation fan 62 is configured to discharge air into the casing 31. During the heat radiation, all the heat radiation fans are turned on, the bottom surface heat radiation fan 62 blows air into the housing 31, and the side surface heat radiation fans 61 on both sides draw air out of the housing 31. Air at room temperature is blown onto the bottom surface of the test tube fixing member 32, and the heat of the test tube fixing member 32 is carried away by the wind and is dissipated by the side cooling fans 61 on both sides, so that a good air cooling effect is achieved. Moreover, since the corresponding side surface of the housing 31 is provided with the corresponding fan heat dissipation hole 311A, even if the heat dissipation fan is removed, the fan heat dissipation hole 311A can also be used as a vent hole, and when the centrifugal operation is performed on the hybrid module 3 through the centrifugal module 2, the external air can enter the housing 31 through the fan heat dissipation hole to play a role in heat dissipation. At this time, the rotational speed of the centrifugal module 2 can be controlled to realize rapid flow of air in the housing 31, thereby achieving a good heat dissipation effect.
In addition to the fan heat dissipation holes 311A, ventilation heat dissipation holes 311B are provided on the housing 31 of the present embodiment to further improve the heat dissipation effect. The housing 31 of the present embodiment is provided with a plurality of ventilation and heat dissipation holes 311B arranged side by side,
the nucleic acid isolation apparatus of the present embodiment further includes a control module 4. As shown in fig. 8, the control module 4 includes a master controller 41 and two slave controllers 42 corresponding to the two hybrid modules 3. The master controller 41 and the slave controller 42 solve the electrical wiring problem using a conductive slip ring 43. As shown in fig. 2, an electrically conductive slip ring 43 is disposed on the elongate shaft 25. Specifically, the conductive slip ring 43 includes a stator end and a rotor end, the stator end is fixed on the motor bracket 28 through a fixing bracket, an electrical signal line of the stator end is connected with the main controller 41, the rotor end is fixed with the extension shaft 25, and an electrical signal line of the rotor end is connected with the slave controller 42, so that the main controller is electrically connected with the slave controller, and transmission of power current and signals can be realized. The conductive slip ring is internally provided with an electric brush and a bearing structure, and is kept in sliding contact with the electric brush when rotating so as to transmit power current and signals. The conductive slip ring structurally adopts high-temperature alloy and composite materials, and cooling is not needed in the use process.
The centrifugal module 2 is controlled by a master controller 41, the mixing module 3 and the heating module 5 are controlled by slave controllers 42, and the two slave controllers 42 are respectively fixed on two sides of the centrifugal tray 21 to respectively control functional modules on the two sides. The main controller 41 is connected with a plurality of groups of buttons and indicator lamps, the buttons are used for giving commands to the main controller 41, the main controller 41 judges the commands, then communicates with the slave controller 422, performs corresponding operations, and controls the indicator lamps to reflect the running states of the device by the main controller 41. The user can use the button operation device and read the operating state of the device by the indicator lamp.
The working procedure of the nucleic acid extraction apparatus of this embodiment is as follows:
the nucleic acid reagent is put into the test tube A, which is put into the test tube receiving hole 321 on the test tube holder 32, and locked by the test tube fastening strip 39. In the mixing process, the output shaft of the stepper motor 35 performs periodic circular motion to drive the shell 31 to rotate, and at the moment, the test tube A placed in the shell 31 performs upside down motion, and the nucleic acid reagent in the test tube A is also upside down so as to achieve the purpose of upside down mixing of the nucleic acid reagent in the test tube A. The heating control is performed on the electrothermal film, so that the reagent in the test tube A is heated. The heating control can be performed simultaneously with the mixing control, i.e. the test tube a is moved upside down while the mixing module 3 performs the mixing operation, and the heating module 5 simultaneously performs the heating control on the reagent.
After the mixing and heating are completed, the reagent needs to be centrifugally separated, the centrifugal module 2 performs centrifugal operation under the condition that the left and right sides of the mixing module 3 are symmetrical, and the mixing module 3 and the heating module 5 which are arranged on the centrifugal tray 21 are driven to be centrifugally separated together, so that the reagent in the test tube A is separated under the action of centrifugal force.
The heating module 5 of this embodiment is mounted in the mixing module 3, both of which are mounted on the centrifugal module 2, so that the reagents can be mixed upside down at an angle on an inclined plane while accompanying heating control, and can also perform high-speed circular motion along the centrifugal axis. Meanwhile, the test tube A is placed at a certain angle, so that the reagent can be separated under the action of centrifugal force. The nucleic acid extraction device of this embodiment integrates three functional modules on one platform, selects combinations of different functions according to the requirements of the nucleic acid extraction flow, and completes the nucleic acid automatic extraction process by cooperation of the modules. Wherein, mixing module and heating module are got up by metal casing protection, prevent that the maloperation from causing the injury to personnel when high temperature heating, high-speed centrifugation.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same; while the invention has been described in detail with reference to the preferred embodiments, those skilled in the art will appreciate that: modifications may be made to the specific embodiments of the present invention or equivalents may be substituted for part of the technical features thereof; without departing from the spirit of the invention, it is intended to cover the scope of the invention as claimed.

Claims (13)

1. A nucleic acid extraction device, comprising:
a base (1);
the centrifugal module (2) comprises a centrifugal tray (21) and a centrifugal driving device for driving the centrifugal tray (21) to rotate, and the centrifugal tray (21) is rotatably arranged on the base (1) around a centrifugal axis; and
mixing module (3), including test tube holder (32), test tube holder (32) have be used for holding test tube (A) test tube accommodation hole (321), mixing module (3) under drive arrangement's drive for centrifugal tray (21) rotate around mixing axis circumference in order to realize mixing to reagent, just test tube holder (32) around mixing axis rotationally set up in centrifugal tray (21), mixing axis for centrifugal axis slope sets up, test tube accommodation hole (321) the axis with mixing axis is crossed, nucleic acid extraction device still includes heating module (5), heating module (5) include heating piece, heating piece pair test tube holder (32) heat, mixing module (3) still including set up in casing (31) in the outside of test tube holder (32), casing (31) include cell body (311) and rotate connect in lid (312) on cell body (311), be provided with fastening strip (39) on the inner wall of lid (312) for centrifugal axis slope sets up, test tube holder (321) have the test tube holder (39) and hold down in the test tube holder (32) in the test tube holder (311) position when holding down test tube holder (39), the test tube fastening strip (39) is provided with a limiting groove, and the limiting groove extends along the distribution direction of the test tube accommodating holes (321) so that the test tube cover is clamped in the limiting groove.
2. The nucleic acid extraction device of claim 1, wherein the mixing axis is inclined at an angle in the range of 15 ° to 45 ° relative to the centrifugal axis.
3. The nucleic acid extraction device according to claim 1, characterized in that the axis of the cuvette receiving hole (321) is perpendicular to the mixing axis.
4. The nucleic acid extraction device according to claim 1, characterized in that the cuvette holder (32) is configured to be periodically positively and negatively rotated with respect to the centrifugation tray (21).
5. The nucleic acid extraction device according to claim 1, characterized in that it comprises two mixing modules (3) arranged symmetrically with respect to the centrifugal axis.
6. The nucleic acid isolation apparatus according to claim 5, wherein the centrifugal tray (21) includes a central tray body (211) and inclined tray bodies (212) provided on both sides of the central tray body (211), and the two mixing modules (3) are respectively provided on the two inclined tray bodies (212) in correspondence.
7. The nucleic acid isolation apparatus according to claim 1, wherein an elastic material is installed in the limit groove.
8. The nucleic acid extraction apparatus according to claim 1, characterized in that the cuvette holder (32) includes a connection side (322) on both sides of the bottom, the mixing module (3) further includes a spacer (33) provided between the connection side (322) and the bottom surface of the housing (31), and the connection side (322) is connected to the housing (31) through the spacer (33).
9. The nucleic acid isolation apparatus according to claim 1, wherein the heating member is an electrothermal film, and the electrothermal film is attached to an end surface of the cuvette holder (32).
10. The nucleic acid extraction apparatus according to claim 1, characterized in that the heating module (5) further comprises a temperature sensor, and the cuvette holder (32) further comprises a detection hole provided between two adjacent cuvette receiving holes (321), the temperature sensor being placed in the detection hole.
11. The nucleic acid extraction apparatus according to any one of claims 1 to 10, further comprising a heat radiation module (6), wherein the heat radiation module (6) comprises a side heat radiation fan (61) provided to a side of the cuvette holder (32); and/or, the heat dissipation module (6) further comprises a bottom surface heat dissipation fan (62) arranged on the bottom surface of the test tube fixing piece (32).
12. The nucleic acid extraction apparatus according to claim 11, characterized in that the mixing module (3) comprises a housing (31), and a fan heat radiation hole is provided in the housing (31).
13. The nucleic acid extraction device according to claim 1, further comprising a control module (4), the control module (4) comprising a main controller (41) coupled to the centrifugation module (2), a secondary controller (42) coupled to the mixing module (3), and an electrically conductive slip ring (43), the electrically conductive slip ring (43) being connected to the centrifugation tray (21) and electrically connecting the main controller (41) and the secondary controller (42).
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CN110437970A (en) * 2019-05-15 2019-11-12 湖北微伞医疗科技有限公司 Intestinal flora nucleic acid extraction system

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CN102102080A (en) * 2010-11-24 2011-06-22 清华大学 Automatic biological component extracting device
CN204111737U (en) * 2014-09-17 2015-01-21 深圳市宝安区沙井人民医院 A kind of high channel nucleic acid extraction purifying many connecting legs eccentric angle rotor
CN105259355A (en) * 2015-09-23 2016-01-20 浙江大学 Automatic biological sample pretreatment device
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