CN110923124A - Instrument for extracting and purifying nucleic acid by magnetic bead method - Google Patents
Instrument for extracting and purifying nucleic acid by magnetic bead method Download PDFInfo
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Abstract
The invention discloses an instrument for extracting and purifying nucleic acid by a magnetic bead method, which comprises a rack, a main shaft, an extraction assembly, a bottom plate and a baffle assembly. Wherein, the extraction component comprises two groups of extraction components which are arranged in parallel in an independent operation mode. The bottom plate sets up a plurality of operation positions, and each operation position sets up four fixed positions of perforated plate respectively. The main shaft is fixed on the frame and is arranged to enable the two groups of extraction assemblies to move between a plurality of operation positions simultaneously or independently. In addition, the shutter can be moved to or from the lower portion of the extraction assembly by motor drive. The instrument of the invention overcomes the problems of poor alignment between the magnetic rod and the magnetic rod sleeve, poor running coordination between arrays and other instrument running stability caused by flux increase, can realize nucleic acid extraction with the flux as high as 384, can also extract a small amount of samples with low cost, and improves the utilization rate of the instrument.
Description
Technical Field
The invention relates to the field of nucleic acid extraction and purification, in particular to an instrument for extracting and purifying nucleic acid by a magnetic bead method.
Background
In biological laboratories, the isolation and purification of biomolecules such as nucleic acids is an important and indispensable task. In a large number of researches, nucleic acid extraction is the first step of subsequent experiments or researches such as molecular diagnostic experiments, and the subsequent researches can be carried out only after nucleic acid is extracted and purified, and the method is also a very important link, and the importance of nucleic acid extraction and purification can be seen.
The traditional nucleic acid extraction and purification methods are not technically difficult, but most of the traditional methods are manual methods, are complex in operation, waste time and labor, and are not suitable for extracting and purifying nucleic acid of a large number of samples. Various magnetic bead-based nucleic acid extraction instruments have been developed, however, these instruments have a relatively small throughput. For example, an IAUTOMAG nucleic acid extractor, 32 channels, manufactured by BioTeke. A nucleic acid extractor, MAGAX 24 manufactured by ABI, is a 24-channel nucleic acid extractor. At present, the largest flux is Kingfisher Flex produced by Sammerfo, the flux is 96, but at most, nucleic acid extraction of 96 samples can be carried out at one time.
Although these current instruments are widely used in various fields such as genomics, disease control, medical treatment, food safety, and forensic identification, the throughput thereof is insufficient to cope with analysis of a large number of samples. Therefore, there is still a need for a nucleic acid extraction and purification apparatus with higher throughput and higher efficiency, so as to meet the requirement for extracting and purifying high-throughput samples.
Disclosure of Invention
In view of at least some of the technical problems in the prior art, the inventors have devised a multi-array, dual-extraction-module instrument that makes it possible to perform nucleic acid extraction up to 384 fluxes, while also performing extraction of a small number of samples at low cost by selectively operating different extraction modules, thereby enabling the instrument to be applied to different situations and improving the utilization of the instrument. In addition, the instrument has a specific structural layout, and the problems of poor alignment between the magnetic rod and the magnetic rod sleeve, poor running coordination between arrays and other instrument running stability caused by flux increase are solved. Specifically, the present invention includes the following.
An instrument for extracting purified nucleic acid by a magnetic bead method comprises a frame, a main shaft, an extraction assembly, a bottom plate and a baffle assembly; wherein:
the extraction assembly comprises a first extraction assembly and a second extraction assembly which are arranged in parallel in an independent operation mode, so that the first extraction assembly and/or the second extraction assembly can be selectively operated according to needs; the first extraction assembly and the second extraction assembly respectively comprise a magnetic bar module, a magnetic bar sleeve module and a fixing module, the magnetic bar module comprises a magnetic bar rack piece, a left magnetic bar array, a right magnetic bar array and a motor for the magnetic bar module, the magnetic bar sleeve module comprises a left magnetic bar sleeve array, a right magnetic bar sleeve array, a magnetic bar sleeve rack piece and a motor for the magnetic bar sleeve module, and the fixing module is used for enabling the extraction assembly to be movably arranged on the main shaft;
the left magnetic rod array and the right magnetic rod array are respectively 12X8 arrays and are respectively fixed on the magnetic rod machine frame piece, and the magnetic rod module is fixed on the magnetic rod sleeve machine frame piece by a motor and is arranged to drive the magnetic rod machine frame piece to move relative to the magnetic rod sleeve module; the left magnetic bar array is matched with the left magnetic bar sleeve array, the right magnetic bar array is matched with the right magnetic bar sleeve array, so that each magnetic bar in the left magnetic bar array and the right magnetic bar array can freely enter and exit the corresponding magnetic bar sleeve, the left magnetic bar sleeve array and the right magnetic bar sleeve array can be respectively combined with or separated from the magnetic bar sleeve frame piece, and the magnetic bar sleeve module is fixed on the fixed module by a motor and is arranged to drive the magnetic bar sleeve frame piece to move relative to the magnetic bar module;
a plurality of operation positions are arranged on the bottom plate along the moving direction of the extraction assembly, each operation position has a size corresponding to the extraction assembly, and four porous plate fixing positions are respectively arranged in each operation position along the direction vertical to the moving direction of the extraction assembly;
the main shaft is fixed on the frame and is arranged to enable the first extraction assembly and the second extraction assembly to move between the plurality of operation positions simultaneously or independently;
the baffle assembly comprises a baffle and a baffle motor, wherein the baffle motor is arranged to drive the baffle to move, so that the baffle can move to the lower part of the extraction assembly or move out of the lower part of the extraction assembly.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, a plurality of limiting devices are arranged on the spindle, each limiting device is used for enabling the first extraction assembly and the second extraction assembly to respectively correspond to different operation positions, the first extraction assembly is movably arranged on the first spindle, and the second extraction assembly is movably arranged on the second spindle.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, the fixing module preferably comprises a first fixing plate, a second fixing plate and aluminum; the first fixing plate is used for being connected with the main shaft, the second fixing plate is used for being connected with the magnetic rod module and the magnetic rod sleeve module, and the first fixing plate and the second fixing plate are connected through the angle aluminum to form an inverted L-shaped structure.
According to the apparatus for extracting purified nucleic acid by the magnetic bead method of the present invention, preferably, the magnetic rod sleeve module is fixed to the first fixing plate and the second fixing plate by a motor through a fixing bracket, and the magnetic rod sleeve module is further fixed to the second fixing plate by a rotating shaft of the motor.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, the second fixing plate is provided with a slide rail, and the magnetic rod rack part and the magnetic rod sleeve rack part are respectively provided with a slide block matched with the slide rail.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, the magnetic rod rack part is arranged above the magnetic rod sleeve rack part, and the slide block of the magnetic rod rack part and the slide block of the magnetic rod sleeve rack part are respectively matched and connected with the slide rail.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, the second fixing plate is provided with a first slide rail and a second slide rail; the magnetic rod rack piece is provided with a first magnetic rod rack piece sliding block and a second magnetic rod rack piece sliding block; the magnetic bar sleeve machine frame part is provided with a first sliding block of the magnetic bar sleeve machine frame part and a second sliding block of the magnetic bar sleeve machine frame part; the first slide block of the magnetic rod rack piece and the first slide block of the magnetic rod sleeve rack piece are respectively matched and connected with the first slide rail, and the second slide block of the magnetic rod rack piece and the second slide block of the magnetic rod sleeve rack piece are matched and connected with the second slide rail.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, the motor for the magnetic rod sleeve module is a 57-step motor, and the motor is also used for driving the magnetic rod module and the magnetic rod sleeve module to vibrate.
The apparatus for extracting and purifying nucleic acid by the magnetic bead method according to the present invention preferably further comprises a heating part disposed below the bottom plate for heating at least a part of the multi-well plate.
According to the instrument for extracting and purifying nucleic acid by the magnetic bead method, preferably, the magnetic rod rack component is provided with two rack blocks corresponding to the two magnetic rod arrays respectively, the two rack blocks form a shape matched with the magnetic rod sleeve rack component, the lower part of the rack block is provided with a shape capable of fixing the magnetic rod arrays, and the upper part of the rack block is provided with a side wing extending upwards.
The instrument of the invention adopts multi-array combination, selectively operates the design of different extraction components, can extract nucleic acid of a small amount of samples at low cost while having a flux as high as 384, improves the utilization rate of the instrument and enriches the application scenes of the instrument.
In the instrument development process, through a series of structural design, especially the cross position layout between bar magnet module and bar magnet cover module motor, motors such as connection between different parts have overcome along with the flux increase, the quantity grow of bar magnet and bar magnet cover in the array, hardly guarantee the smooth complex problem of every bar magnet and bar magnet cover accurately, guarantee the effective cooperation of each bar magnet and bar magnet cover in different arrays, and then guaranteed the steady operation of instrument. In addition, possible contamination problems between different samples due to increased throughput are avoided by designing the baffle assembly.
The instrument has the characteristics of simple operation, large flux and high efficiency, and has wide application prospect in the field of high-flux sample extraction and purification.
Drawings
FIG. 1 is a schematic diagram showing an exemplary structure of an apparatus for extracting purified nucleic acid by a magnetic bead method.
Fig. 2 is a schematic structural view of the remaining part of fig. 1 with the protective casing omitted.
Fig. 3 is a schematic diagram illustrating an instrument structure of two extraction assemblies.
FIG. 4 is a schematic diagram of an exemplary extraction assembly of an apparatus for magnetic bead extraction of purified nucleic acids.
FIG. 5 is a schematic diagram of an extraction module from another perspective of an apparatus for extracting purified nucleic acids by magnetic bead method.
FIG. 6 shows the structure of an exemplary immobilization module of an apparatus for extracting purified nucleic acid by the magnetic bead method and the relationship of immobilization.
FIG. 7 is an explanatory view from another angle of an apparatus for extracting purified nucleic acid by magnetic bead method, in which the structures of a fixing module and a baffle are shown.
FIG. 8 is a schematic diagram of an exemplary slide and slide track of an apparatus for extracting purified nucleic acids by magnetic bead method.
FIG. 9 is a schematic diagram of an exemplary bottom plate of an apparatus for extracting purified nucleic acid by a magnetic bead method.
Fig. 10 is a diagram exemplarily showing a bar magnet array and a bar magnet sleeve array and their mating relationship.
Description of reference numerals:
100 … rack; 200 … a main shaft; 110 … protective housing; 111 … case front door; 112 … display panel; 300 … extraction component; 301 … a first extraction assembly; 302 … second extraction component; 310 … magnetic bar module; 320 … a magnet bar sleeve module; 360 … slide rails; 311 … bar magnet frame pieces; 3111 … frame blocks; 312 … magnetic bar array; 313 … motor for magnetic bar sleeve module; 321 … magnetic rod set frame parts; 3211 … side wing; 322 … array of magnetic rod sleeves; 323 … motor for magnetic bar module; 341 … first retainer plate, 342 … angle aluminum; 343 … second fixing plate; 351 … bar magnet frame member first slide block; 353 … magnetic bar sleeve frame member first slide block; 400 … baffle plate; 401 … baffle motor; 500 … a base plate; 510 … perforated plate fixing position; 520, 520 … perforated plates; 600 … heating section.
Detailed Description
Various exemplary embodiments of the present invention are described in detail below with reference to the accompanying drawings, which should not be considered limiting of the invention, but rather are understood to be more detailed descriptions of certain aspects, features and embodiments of the invention.
The terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under particular conditions and that the embodiments described herein are capable of operation in other sequences than described or illustrated herein. Also, the terms left, right, top, bottom, over, under and the like in the description and the claims are used for descriptive purposes and not necessarily for describing relative positions.
The instrument for extracting and purifying nucleic acid by the magnetic bead method is an instrument for separating (also called extracting) and purifying nucleic acid in the field of biochemistry, and adopts a multi-array and double-extraction-component design. The device mainly comprises a rack, a main shaft, an extraction assembly, a bottom plate and a baffle plate assembly. Wherein, the extraction element is two sets of cophase assemblies, respectively the first extraction element and the second extraction element. The components are described in detail below.
[ frame ]
The rack of the invention is mainly used for fixing the protective shell so as to protect internal components of the instrument, such as the extraction assembly, the main shaft, the bottom plate and the like, the protective shell can be arranged outside the rack, the connection mode of the rack and the protective shell is not particularly limited, for example, the rack and the protective shell are fixedly connected through a groove clamp or a nut, the rack and the protective shell form a box body, the shape of the box body is not particularly limited, for example, the box body can be a cuboid, a cube or an irregular columnar structure. In a specific embodiment, the extraction and purification instrument of the invention is a box body with a cuboid structure. Preferably, one end of the protective shell is provided with a shell front door, the protective shell can further comprise a switch, and the bottom of the protective shell is an incompletely-closed bottom shell.
[ extraction Assembly ]
The extraction components of the invention are divided into two groups, which form the main part of nucleic acid extraction. Each extraction assembly comprises a magnetic rod module, a magnetic rod sleeve module and a fixing module. The magnetic rod module and the magnetic rod sleeve module are respectively composed of a plurality of 2 arrays for example. Although the design of adopting multiple arrays and double extraction assemblies can simultaneously carry out high-throughput and low-energy nucleic acid extraction, the design also easily causes the problem that the magnetic rod and the magnetic rod sleeve are not positioned accurately, so that the instrument is poor in stability. The reason for this is that, firstly, the diameter of the magnetic rod and the magnetic rod sleeve is only a few millimeters, and it is difficult to ensure accurate synchronous coordinated operation between different arrays in a high-throughput instrument using multiple arrays, so as to ensure that each magnetic rod with such a small diameter can smoothly enter and exit the magnetic rod sleeve at the same time. Secondly, for the magnetic bead method extraction instrument, after the magnetic rod enters the magnetic rod sleeve, the process that the assembly consisting of the magnetic rod and the magnetic rod sleeve vibrates in the test tube to ensure that the sample is fully mixed is involved, and the vibration is the key for ensuring the extraction purity of the nucleic acid, but the vibration is very unfavorable for the matching between the magnetic rod and the magnetic rod sleeve in the array.
In order to solve the problems, the extraction assembly provided by the invention adopts a mode that the magnetic rod modules and the magnetic rod sleeve modules are arranged in a crossed manner and are further connected with the fixing module through sliding connection, so that the positioning accuracy and stability of each magnetic rod in a multi-array are improved. The structure of the extraction assembly is described in detail below.
The bar magnet module of the extraction assembly of the present invention generally comprises a bar magnet frame member, a bar magnet array and a motor for the bar magnet module. The magnetic bar array is used for adsorbing or transferring magnetic beads and adopts a 12X8 specification. The magnetic bar module of the present invention comprises two magnetic bar arrays. Referred to herein as a left bar magnet array and a right bar magnet array. The magnetic rod array of the invention comprises 96 magnetic rods which are vertically arranged. The magnetic rods can be fixed on the fixing plate and fixed on the magnetic rod machine frame component through the fixing plate.
The bar magnet frame member of the present invention is used for fixing the bar magnet array, and the shape and size thereof are not particularly limited. In certain embodiments, the bar magnet frame pieces have a shape adapted to secure (e.g., snap fit) the left and right bar magnet arrays side-by-side. The bar magnet module is secured to the bar magnet housing frame member with a motor and is configured to drive the bar magnet frame member to move, e.g., move up and down, relative to the bar magnet housing module. This arrangement is advantageous in keeping each bar magnet in both arrays moving smoothly to ensure accurate positioning. In the invention, a motor is used for driving the bar magnet frame piece in the first extraction module to move relative to the bar magnet sleeve module. Similarly, another motor is used to drive the bar magnet frame member in the second extraction module to move relative to the bar magnet nest module.
In the invention, the magnetic bar sleeve module generally comprises a magnetic bar sleeve machine frame component, a magnetic bar sleeve array and a motor for the magnetic bar sleeve module. The magnetic rod sleeve array comprises a left magnetic rod sleeve array and a right magnetic rod sleeve array, wherein the left magnetic rod sleeve array is matched with the left magnetic rod array, and the right magnetic rod sleeve array is matched with the right magnetic rod array, so that each magnetic rod of the magnetic rod array can freely enter and exit the corresponding magnetic rod sleeve of the magnetic rod sleeve array. The left magnetic rod sleeve array and the right magnetic rod sleeve array are respectively provided with 12X8 magnetic rod sleeves. Each magnetic rod sleeve is matched with a magnetic rod and is in a shape that the magnetic rod sleeve can freely enter and exit. In certain embodiments, the magnetic rod sleeve of the present invention is a plastic tube and the magnetic rod is free to enter or exit the lumen of the plastic tube.
In the invention, the motor for the magnetic bar sleeve module is used for driving the magnetic bar sleeve frame piece to move up and down relative to the magnetic bar module. Preferably, the device can be further used to drive the oscillation of the extraction assembly under program control to achieve mixing of the liquids. In order to reduce the influence of vibration on the stability of the instrument or the positioning of the magnetic rod, the magnetic rod is sleeved by the motor and is fixed on the first fixing plate and the second fixing plate of the fixing module through the fixing bracket. To enhance the stability, the rotating shaft of the motor is further fixed to the second fixing plate by a fixing position.
[ baseboard ]
The base plate of the present invention is used to support a plurality of operating positions. Generally, a plurality of operation bits, for example, 5 operation bits are sequentially set in the moving direction along the extracting member. Herein, a manipulation site refers to a spatial location for completing a certain process or step of nucleic acid extraction (e.g., sample mixing, DNA binding, washing, elution, etc.). The operation position is provided with four perforated plate fixing positions, and each fixing position can fix one 12X8 perforated plate. A corresponding number of multi-well plates may be placed during operation of the instrument depending on the number of magnetic bar arrays. The multi-well plate may be fixed at a corresponding multi-well plate fixing position before the nucleic acid extraction work is started. The multiwell plate may be pre-filled with a corresponding reagent or liquid, so that the process can be automated according to a predetermined procedure or a program preset in the apparatus. According to the invention, the arrangement mode of the multiple operation positions and the multi-plate fixing positions ensures that the same type of reagent or sample is placed in each hole in each multi-plate during operation, and compared with the mode of placing different types of reagents in one multi-plate, the multi-plate multi-hole reagent placing device is convenient to operate and higher in efficiency.
The apparatus for extracting and purifying nucleic acid by the magnetic bead method of the present invention may preferably further comprise a heating unit disposed below the bottom plate for heating at least a part of the multi-well plate. The heating part and the heating method thereof are not particularly limited as long as the extracted DNA molecules are heated by heating the multi-well plate, and in a specific embodiment, the pre-heating is performed to 65 ℃.
[ Main shaft ]
The main shaft of the invention is fixed on the frame and is arranged to enable the first extraction assembly and the second extraction assembly to move between a plurality of operation positions simultaneously or independently. Preferably, the main shaft is provided with limit grooves corresponding to different operation positions, so that the extraction assembly is favorably fixed above the corresponding operation positions. The movement herein may be any manner suitable for the extraction component to be transferred between different operating positions. For example by means of a drive and a slide. The driving means is not particularly limited, and those known in the art can be used. For example, the drive may be a hydraulic cylinder, an electric motor, or an electric motor, preferably an electric motor drive, such as a 42-step motor. The sliding device is not particularly limited, and may include, for example, a linear guide, a guide bush, a crawler drive, or a sliding screw for movement. The number of the main shafts can be one or two. For example, when the spindle includes a first spindle and a second spindle, the first extraction assembly is movably connected with the first spindle through the fixing module, and the second extraction assembly is movably connected with the second spindle through the fixing module.
[ fixed Module ]
The fixing module comprises fixing plates for fixing different parts, and comprises a first fixing plate, a second fixing plate and angle aluminum. The first fixing plate is used for being connected with the main shaft, and the second fixing plate is used for connecting or fixing the magnetic rod module and the magnetic rod sleeve module. The first fixing plate and the second fixing plate are connected through angle aluminum to form an inverted L-shaped structure. The shape and number of the fixing plates of the present invention are not particularly limited. The number of the second fixing plates may be determined according to the number of the extraction members.
The second fixing plate of the fixing module can be provided with a slide rail, so that the magnetic rod sleeve frame piece and the magnetic rod frame piece can be arranged on the fixing module in a sliding mode through the slide block. In some embodiments, a slide rail is vertically disposed on the second fixing plate, and the magnetic rod rack member is provided with a slide block which is matched with the slide rail. Similarly, the magnetic bar sleeve frame part is provided with a slide block matched with the slide rail, and the slide block of the magnetic bar sleeve frame part share the same slide rail.
In an exemplary embodiment, the second fixing plate is provided with two slide rails, the first slide rail and the second slide rail are arranged in parallel on one side of the second fixing plate facing the inside of the rack in a vertical manner, meanwhile, the magnetic bar rack part is provided with a magnetic bar rack part first slide block and a magnetic bar rack part second slide block, the magnetic bar sleeve rack part is provided with a magnetic bar sleeve rack part first slide block and a magnetic bar sleeve rack part second slide block, and the magnetic bar rack part first slide block and the magnetic bar sleeve rack part first slide block share the first slide rail, namely, the magnetic bar rack part first slide block and the magnetic bar sleeve rack part first slide block are respectively and slidably arranged on the first slide rail; meanwhile, the second slide block of the magnetic rod rack piece and the second slide block of the magnetic rod sleeve rack piece share a second slide rail, namely the second slide block of the magnetic rod rack piece and the second slide block of the magnetic rod sleeve rack piece are respectively arranged on the second slide rail in a sliding manner.
[ baffle plate Assembly ]
The baffle plate assembly comprises a baffle plate and a motor for the baffle plate. The baffle plate component is arranged to prevent the liquid in different holes from being polluted when the extraction component moves. This is important for the throughput sample processing of the present invention.
In the invention, the baffle is driven to move by the motor, so that the baffle can move to the lower part of the extraction assembly or move out of the lower part of the extraction assembly, and the effect of cross contamination caused by liquid dropping from the previous operation position to other operation positions in the operation process is prevented. The number of baffles can be determined according to the number of extraction assemblies and also according to the number of arrays. Preferably, the first extraction group is provided with 1 baffle and motor for the baffle, the second extraction group is provided with 1 baffle and motor for the baffle, and the baffle and motor for the baffle of the first extraction group and the baffle and motor for the baffle of the second extraction group are arranged in parallel.
Examples
The structure of an exemplary instrument of the present invention is described below with reference to the accompanying drawings. For convenience of explanation, some components may be omitted in the following drawings. These drawings are for illustrative purposes only and should not be used to narrowly interpret the scope of the claimed invention.
FIG. 1 is a schematic external view of an apparatus for extracting purified nucleic acid by the magnetic bead method. As shown in FIG. 1, a protective casing 110 is provided outside a rack 100, and the rack of the apparatus for extracting and purifying nucleic acid by magnetic bead method of the present invention is mainly used for fixing the protective casing and a spindle and a base plate described below, and the rack is fixedly connected with the protective casing to form a box body with a rectangular structure. A front housing door 111 is disposed at one end of the protective housing 110, a display panel 112 is disposed at one side of the front housing door, the protective housing 110 further includes a switch (not shown in the figure), and the bottom of the protective housing 110 is an incompletely closed bottom housing.
Fig. 2 is a schematic structural view of fig. 1 with the protective casing omitted. As shown in FIG. 2, the apparatus for extracting purified nucleic acid by the magnetic bead method of this embodiment 1 includes a housing 100, a spindle 200, an extraction module 300, and a base plate 500. The instrument of this embodiment comprises 5 operating positions. Four perforated plates are placed on the perforated plate fixing positions of each operation position. The two spindles 200 of the present invention are respectively fixed to the upper portion of the inside of the frame 100. A motor, such as a 42 stepper motor, is provided at one end of the spindle 200. The main shaft is provided with a slide block. With the above-described structure, the extracting assembly 300 is horizontally moved along the main shaft 200 inside the instrument with respect to the housing 100.
FIGS. 3, 4 and 5 schematically show diagrams of an extraction assembly of an instrument for magnetic bead extraction of purified nucleic acids. As shown, the extraction assembly 300 of the present embodiment includes a first extraction assembly 301 and a second extraction assembly 302 arranged in parallel. The two extraction assemblies are slidably connected to the two spindles 200 by a fixed plate. The two extraction assemblies have the same structure and composition.
In the apparatus shown in fig. 5, the right-hand extraction module is used as the first extraction module, and the left-hand extraction module is used as the second extraction module. Each extraction assembly of the present embodiment includes a bar magnet module 310, a bar magnet sleeve module 320, and a fixed module. The magnetic rod module comprises a magnetic rod frame piece 311, a magnetic rod array 312 and a motor 323 for the magnetic rod module. The rod cover module 320 includes a rod cover frame member 321, a rod cover array 322, and a rod cover module motor 313. In this embodiment, the bar magnet array 312 includes a left bar magnet array and a right bar magnet array. Taking fig. 5 as an example, the left magnetic bar array and the right magnetic bar array are two magnetic bar arrays located at the rightmost side of fig. 5. The second extraction assembly has the same structure as the first extraction assembly. The bar magnet array 312 is composed of 96 bar magnets of 12X8 vertical, and is fixed on the bar magnet frame 311. The left magnetic bar array and the right magnetic bar array are fixedly arranged on the magnetic bar frame piece 311 in parallel.
Fig. 6 shows the structure and connection of the bar magnet frame pieces and the bar magnet sleeve frame pieces. As shown in fig. 6, the bar magnet frame assembly 311 has a structure that can be sleeved inside the bar magnet frame assembly 321, and each bar magnet frame assembly 311 has two frame blocks 3111 corresponding to two bar magnet arrays, respectively, and the two frame blocks 3111 form a shape matching with one bar magnet frame assembly 321. Each rack block 3111 has a shape capable of fixing a magnetic bar array on a lower side thereof, and has a side wing extending upward on an upper side of the rack block 3111. The two frame blocks 3111 are fixedly connected to the slider through an angle aluminum provided in the horizontal direction. Similarly, there are upwardly extending wings 3211 on both sides of the bar set frame member 321, the wings 3211 advantageously prevent contamination of liquid between the different arrays that may occur from splashing. Similarly, the rod housing 321 is fixedly connected to the slider by means of an aluminum member disposed horizontally.
In this embodiment, the bar magnet frame 311 and the bar magnet sleeve frame 321 are disposed in an up-down manner, and the bar magnet array 312 of the bar magnet frame 311 and the bar magnet sleeve array 322 of the bar magnet sleeve frame 321 are disposed correspondingly, so that each bar magnet can freely enter and exit the sleeve inner cavity of the corresponding bar magnet sleeve along with the relative movement of the bar magnet module and the bar magnet sleeve module. The design is beneficial to improving the positioning accuracy of the magnetic rod.
In this embodiment, the motor 323 for the magnetic rod module is fixed below the horizontal fixing plate of the magnetic rod sleeve frame 321, and the screw rod of the motor 323 for the magnetic rod module sequentially passes through the horizontal fixing plate of the magnetic rod sleeve frame 321 and the horizontal fixing plate of the magnetic rod frame 311 upward and is connected to the screw rod cap fixed to the magnetic rod frame 311. This configuration allows the motor to drive the movement of the rod housing piece 311 relative to the rod sleeve module 320. In this embodiment, each extraction module has a motor for a bar magnet module. I.e. two bar magnet modules share one motor.
The magnetic rod sleeve array of this embodiment includes two magnetic rod sleeve arrays. Taking fig. 5 as an example, the left and right bar magnet sleeve arrays in the first extraction assembly on the right side are not shown. The second extraction assembly on the left side comprises a third magnetic rod sleeve array and a fourth magnetic rod sleeve array, the third magnetic rod sleeve array is matched with the third magnetic rod array, and the fourth magnetic rod sleeve array is matched with the fourth magnetic rod array. The magnetic rod sleeve array is matched with the magnetic rod array, and the magnetic rod sleeve array can be combined with or separated from the magnetic rod sleeve frame piece.
In this embodiment, the motor 313 for the bar magnet cover module is fixed to one side of the second fixing plate by a motor fixing bracket. The lead screw of the motor 313 passes through the horizontal plate of the magnetic bar rack 311 and the horizontal plate of the perforated plate rack 321 in turn downward, and is connected with a lead screw cap fixed below the horizontal plate of the perforated plate rack 321. With this configuration, the motor 313 can drive the rod housing frame 321 to move up and down with respect to the rod module 310, while ensuring stability of movement. In this embodiment, each extraction module uses a motor 313 for a bar magnet cover module.
FIG. 7 schematically shows the structure of the immobilization module of the apparatus for extracting purified nucleic acid by the magnetic bead method and the relationship of the immobilization connection thereof. As shown in fig. 7, the fixing module includes a first fixing plate 341, a second fixing plate 343, and an angle aluminum 342 for fixedly connecting the first fixing plate 341 and the second fixing plate 343, thereby forming an inverted "L" type structure. As shown in fig. 7, the first fixing plate 341 is a horizontal fixing plate, and the second fixing plate 343 is a vertical fixing plate. A slider is provided above the first fixing plate and is mounted to the main shaft 200 through the slider.
Also shown in fig. 7 is an exemplary baffle 400 and baffle motor 401 configuration. As shown in fig. 7, the baffle 400 includes a horizontal plate movable to below the magnetic rod housing or the magnetic rod, a slider for slidably connecting with the motor driving rod, and a connecting rod connecting the horizontal plate to the slider. The slide block can move horizontally along with the driving of the motor, thereby controlling the horizontal movement of the baffle. The first extraction group is provided with 1 baffle and a motor for the baffle, the second extraction group is provided with 1 baffle and a motor for the baffle, and the baffle and the motor for the baffle of the first extraction group and the baffle and the motor for the baffle of the second extraction group are arranged in parallel.
FIG. 8 is a schematic diagram of a slide block and a slide rail structure of an apparatus for extracting purified nucleic acid by a magnetic bead method. The slide rails and the sliders are used as members for moving the bar magnet frame piece 311 and the bar magnet sleeve frame piece 321 up and down. As shown in fig. 8, a slide rail 360 is provided on a side of the second fixing plate 343 facing the inside of the housing. Although only one slide rail 360 is shown in fig. 8, in practice there may be two slide rails 360. A slide block 351 matched with the slide rail 360 is fixed on the magnetic rod frame piece 311. The number of sliders 351 corresponds to the number of slide rails 360. A slider 353 matched with the slide rail 360 is fixed to the bar magnet housing frame member 321 below the bar magnet frame member 311. The number of the sliders 353 corresponds to the number of the slide rails 360. The slider 351 of the rod magnet frame member 311 shares the same slide rail 360 as the slider 353 of the rod magnet frame member 321.
FIG. 9 is a schematic diagram showing the structure of the bottom plate of the apparatus for extracting purified nucleic acid by the magnetic bead method. The bottom plate 500 of the apparatus for extracting purified nucleic acid by the magnetic bead method of the present invention is disposed at the bottom of the rack, and is provided with a multi-well plate fixing station 510 for placing 5 multi-well plates 520. The multi-well plate was a 96-well plate set in a 12X8 format. On the bottom plate 500, 5 operation positions are provided, in the left-to-right direction of the figure, a first operation position, a second operation position, a third operation position, a fourth operation position, and a fifth operation position, which correspond to binding, washing 1, washing 2, washing 3, and elution of nucleic acids, respectively. Each station has four multi-well plate holding stations 510. The apparatus of this embodiment further comprises a heating part 600 disposed below the base plate 500 for heating the multi-well plate of the fifth operation position to pre-heat to 65 c in the previous stage.
Fig. 10 schematically shows a diagram of a magnetic rod array 312 and a magnetic rod sleeve array 322. As shown in fig. 10, the magnetic bar array 312 is composed of 96 magnetic bars vertically arranged on the fixing plate. Accordingly, the magnetic rod sleeve array is composed of 96 plastic tubes vertically arranged on the fixing plate. When the array of magnetic rods 312 is positioned above the array of magnetic rod sleeves and moved downward, each magnetic rod may enter a corresponding plastic tube, respectively.
While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. Many modifications and variations may be made to the exemplary embodiments of the present description without departing from the scope or spirit of the present invention. The scope of the claims is to be accorded the broadest interpretation so as to encompass all modifications and equivalent structures and functions.
Claims (10)
1. An instrument for extracting purified nucleic acid by a magnetic bead method is characterized by comprising a rack, a main shaft, an extraction assembly, a bottom plate and a baffle assembly; wherein:
the extraction assembly comprises a first extraction assembly and a second extraction assembly which are arranged in parallel in an independent operation mode, so that the first extraction assembly and/or the second extraction assembly can be selectively operated according to needs; the first extraction assembly and the second extraction assembly respectively comprise a fixing module, a magnetic bar module and a magnetic bar sleeve module, the magnetic bar module and the magnetic bar sleeve module are sequentially arranged from top to bottom, the magnetic bar module comprises a magnetic bar frame piece, a left magnetic bar array, a right magnetic bar array and a motor for the magnetic bar module, the magnetic bar sleeve module comprises a left magnetic bar sleeve array, a right magnetic bar sleeve array, a magnetic bar sleeve frame piece and a motor for the magnetic bar sleeve module, and the fixing module is used for enabling the extraction assembly to be movably arranged on the main shaft;
the left magnetic rod array and the right magnetic rod array are respectively 12X8 arrays and are respectively fixed on the magnetic rod frame piece, the magnetic rod module is fixed below the magnetic rod sleeve frame piece by a motor, and the motor shaft of the magnetic rod module penetrates through the magnetic rod sleeve module upwards to be connected to the magnetic rod module and is arranged to drive the magnetic rod frame piece to move relative to the magnetic rod sleeve module;
the left magnetic rod array is matched with the left magnetic rod sleeve array, the right magnetic rod array is matched with the right magnetic rod sleeve array, so that each magnetic rod in the left magnetic rod array and the right magnetic rod array can freely enter and exit the corresponding magnetic rod sleeve, the left magnetic rod sleeve array and the right magnetic rod sleeve array can be respectively combined with or separated from the magnetic rod sleeve machine frame component, the magnetic rod sleeve module is fixed on the upper part of the fixed module by a motor, and the motor shaft of the magnetic rod sleeve module downwards penetrates through the magnetic rod module to be connected to the magnetic rod sleeve module and is arranged to drive the magnetic rod sleeve machine frame component to move relative to the magnetic rod module;
a plurality of operation positions are arranged on the bottom plate along the moving direction of the extraction assembly, each operation position has a size corresponding to the extraction assembly, four porous plate fixing positions are respectively arranged in each operation position along the direction vertical to the moving direction of the extraction assembly, and the porous plates have shapes corresponding to the left magnetic rod sleeve array or the right magnetic rod sleeve array;
the main shaft is fixed on the frame and is arranged to enable the first extraction assembly and the second extraction assembly to move between the plurality of operation positions simultaneously or independently;
the baffle assembly comprises a baffle and a baffle motor, wherein the baffle motor is arranged to drive the baffle to move, so that the baffle can move to the lower part of the extraction assembly or move out of the lower part of the extraction assembly.
2. The apparatus of claim 1, wherein the spindle is provided with a plurality of limiting devices, each limiting device is used for enabling the first extraction module and the second extraction module to correspond to different operation positions, and the first extraction module is movably arranged on the first spindle and the second extraction module is movably arranged on the second spindle.
3. The apparatus of claim 1, wherein the fixing module comprises a first fixing plate, a second fixing plate and aluminum; the first fixing plate is used for being connected with the main shaft, the second fixing plate is used for being connected with the magnetic rod module and the magnetic rod sleeve module, and the first fixing plate and the second fixing plate are connected through the angle aluminum to form an inverted L-shaped structure.
4. The apparatus of claim 1, wherein the magnetic rod sleeve module is fixed to the first fixing plate and the second fixing plate by a motor through a fixing bracket, and the magnetic rod sleeve module is further fixed to the second fixing plate by a rotating shaft of the motor.
5. The apparatus of claim 3, wherein the second fixing plate is provided with a slide rail, and the magnetic rod rack member and the magnetic rod sleeve rack member are respectively provided with a slide block matching with the slide rail.
6. The apparatus of claim 5, wherein the magnetic rod rack is disposed above the magnetic rod set rack, and the slide block of the magnetic rod set rack are respectively connected to the slide rail in a matching manner.
7. The apparatus of claim 6, wherein the second fixing plate is provided with a first slide rail and a second slide rail; the magnetic rod rack piece is provided with a first magnetic rod rack piece sliding block and a second magnetic rod rack piece sliding block; the magnetic bar sleeve machine frame part is provided with a first sliding block of the magnetic bar sleeve machine frame part and a second sliding block of the magnetic bar sleeve machine frame part; the first slide block of the magnetic rod rack piece and the first slide block of the magnetic rod sleeve rack piece are respectively matched and connected with the first slide rail, and the second slide block of the magnetic rod rack piece and the second slide block of the magnetic rod sleeve rack piece are matched and connected with the second slide rail.
8. The apparatus of claim 1, wherein the motor for the magnetic rod sleeve module is a 57-step motor, and the motor is further configured to drive the magnetic rod module and the magnetic rod sleeve module to vibrate.
9. The apparatus according to claim 1, further comprising a heating unit disposed under the bottom plate for heating at least a part of the multi-well plate.
10. The apparatus of claim 1, wherein the magnetic rod holder has two holder blocks corresponding to the two magnetic rod arrays, the two holder blocks are formed in a shape corresponding to a magnetic rod set holder, the holder blocks have a shape capable of fixing the magnetic rod arrays below the holder blocks, and the holder blocks have upwardly extending wings at upper portions of the holder blocks.
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| CN202010095202.4A CN110923124B (en) | 2020-02-17 | 2020-02-17 | Instrument for extracting and purifying nucleic acid by magnetic bead method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022104224A1 (en) * | 2020-11-15 | 2022-05-19 | Life Technologies Corporation | System and method for polynucleotide purification |
| CN115678744A (en) * | 2022-11-03 | 2023-02-03 | 广州湾区生物科技有限公司 | Nucleic acid extraction instrument |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030082565A1 (en) * | 2001-10-31 | 2003-05-01 | Jang Gi Young | Method of manufacturing kit for isolating nucleic acids or biological materials, kit manufactured by the method, and apparatus using the kit |
| US20150299766A1 (en) * | 2012-02-23 | 2015-10-22 | Dow Agrosciences Llc | Automation of barrier-based plant nucleic acid and protein extraction |
| CN106318847A (en) * | 2016-08-22 | 2017-01-11 | 杭州奥盛仪器有限公司 | Nucleic acid extraction equipment capable of eccentric oscillation |
| CN106867883A (en) * | 2017-03-03 | 2017-06-20 | 苏州工业园区铂纳医学科技有限公司 | A kind of automatic instrument for extracting nucleic acid |
| CN106916739A (en) * | 2017-05-04 | 2017-07-04 | 广州和实生物技术有限公司 | A kind of single group of extracting is knitted sample nucleic acid and surveys the automatic extracting instrument of OD values |
| CN108441423A (en) * | 2018-04-28 | 2018-08-24 | 绍兴迅敏康生物科技有限公司 | Instrument for extracting nucleic acid |
| WO2019004636A2 (en) * | 2017-06-28 | 2019-01-03 | (주)오상헬스케어 | Automated nucleic acid separation apparatus |
| CN110093343A (en) * | 2019-05-08 | 2019-08-06 | 中国人民解放军军事科学院军事医学研究院 | A kind of integrated portable nucleic acid-extracting apparatus and method for extracting nucleic acid |
-
2020
- 2020-02-17 CN CN202010095202.4A patent/CN110923124B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030082565A1 (en) * | 2001-10-31 | 2003-05-01 | Jang Gi Young | Method of manufacturing kit for isolating nucleic acids or biological materials, kit manufactured by the method, and apparatus using the kit |
| US20150299766A1 (en) * | 2012-02-23 | 2015-10-22 | Dow Agrosciences Llc | Automation of barrier-based plant nucleic acid and protein extraction |
| CN106318847A (en) * | 2016-08-22 | 2017-01-11 | 杭州奥盛仪器有限公司 | Nucleic acid extraction equipment capable of eccentric oscillation |
| CN106867883A (en) * | 2017-03-03 | 2017-06-20 | 苏州工业园区铂纳医学科技有限公司 | A kind of automatic instrument for extracting nucleic acid |
| CN106916739A (en) * | 2017-05-04 | 2017-07-04 | 广州和实生物技术有限公司 | A kind of single group of extracting is knitted sample nucleic acid and surveys the automatic extracting instrument of OD values |
| WO2019004636A2 (en) * | 2017-06-28 | 2019-01-03 | (주)오상헬스케어 | Automated nucleic acid separation apparatus |
| CN108441423A (en) * | 2018-04-28 | 2018-08-24 | 绍兴迅敏康生物科技有限公司 | Instrument for extracting nucleic acid |
| CN110093343A (en) * | 2019-05-08 | 2019-08-06 | 中国人民解放军军事科学院军事医学研究院 | A kind of integrated portable nucleic acid-extracting apparatus and method for extracting nucleic acid |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022104224A1 (en) * | 2020-11-15 | 2022-05-19 | Life Technologies Corporation | System and method for polynucleotide purification |
| CN115678744A (en) * | 2022-11-03 | 2023-02-03 | 广州湾区生物科技有限公司 | Nucleic acid extraction instrument |
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