CN111440703A - Double-extraction-mode nucleic acid extraction workstation and nucleic acid extraction method - Google Patents
Double-extraction-mode nucleic acid extraction workstation and nucleic acid extraction method Download PDFInfo
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- CN111440703A CN111440703A CN202010424015.6A CN202010424015A CN111440703A CN 111440703 A CN111440703 A CN 111440703A CN 202010424015 A CN202010424015 A CN 202010424015A CN 111440703 A CN111440703 A CN 111440703A
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- 238000000605 extraction Methods 0.000 title claims abstract description 78
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 55
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 55
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 55
- 239000007788 liquid Substances 0.000 claims abstract description 49
- 238000007400 DNA extraction Methods 0.000 claims abstract description 34
- 239000011324 bead Substances 0.000 claims abstract description 34
- 238000003860 storage Methods 0.000 claims abstract description 28
- 238000004140 cleaning Methods 0.000 claims abstract description 27
- 238000010828 elution Methods 0.000 claims abstract description 22
- 238000005336 cracking Methods 0.000 claims abstract description 21
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 14
- 230000009089 cytolysis Effects 0.000 claims description 46
- 230000009977 dual effect Effects 0.000 claims description 17
- 238000010438 heat treatment Methods 0.000 claims description 15
- 239000003480 eluent Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000005057 refrigeration Methods 0.000 claims description 5
- 238000003776 cleavage reaction Methods 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 4
- 230000007017 scission Effects 0.000 claims description 4
- 238000010276 construction Methods 0.000 claims 1
- 238000005406 washing Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
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Abstract
The invention relates to the technical field of nucleic acid extraction, in particular to a double-extraction-mode nucleic acid extraction workstation and a nucleic acid extraction method, wherein the double-extraction-mode nucleic acid extraction workstation comprises a workstation, wherein a liquid storage tank and a plate position are arranged on the workstation, the liquid storage tank is used for placing reagents required by DNA extraction, and the plate position is used for placing a cracking plate, a combination plate, a cleaning plate and an elution plate; the centrifugal device is used for separating different products in the sample which is well cracked in the cracking plate under the centrifugal action; the magnetic bar device is arranged on the workbench and used for adsorbing magnetic beads used in DNA extraction; and the mechanical arm device is arranged on the workbench, can add the reagent in the liquid storage tank into the cracking plate, and can shift the cracking plate, the combination plate or the cleaning plate. The invention can reduce the extraction difficulty of nucleic acid and ensure the smooth extraction of nucleic acid.
Description
Technical Field
The invention relates to the technical field of nucleic acid extraction, in particular to a double-extraction-mode nucleic acid extraction workstation and a nucleic acid extraction method.
Background
In the biological field, it is often necessary to extract nucleic acids from a sample and then perform an assay.
In the prior art, an experimenter generally puts a sample into a lysis plate, then adds lysis solution, centrifuges the sample after lysis, then adds the liquid after lysis into a combination plate, and extracts nucleic acid.
Therefore, a double-extraction mode nucleic acid extraction workstation and a nucleic acid extraction method are needed to solve the above technical problems.
Disclosure of Invention
The invention aims to provide a double-extraction-mode nucleic acid extraction workstation and a nucleic acid extraction method, which can reduce the extraction difficulty of nucleic acid and ensure the smooth extraction of nucleic acid.
In order to achieve the purpose, the invention adopts the following technical scheme:
a dual extraction mode nucleic acid extraction workstation comprising:
the device comprises a workbench, wherein a liquid storage tank and a plate position are arranged on the workbench, the liquid storage tank is used for placing reagents required by DNA extraction, and the plate position is used for placing a cracking plate, a combination plate, a cleaning plate and an elution plate;
the centrifugal device is used for separating different products in the sample which is well cracked in the cracking plate under the centrifugal action;
the magnetic bar device is arranged on the workbench and used for adsorbing magnetic beads used in DNA extraction;
and the mechanical arm device is arranged on the workbench, can add the reagent in the liquid storage tank into the cracking plate, and can shift the cracking plate, the combination plate or the cleaning plate.
Optionally, the robot arm device includes a robot arm, a liquid transferring pump, and a gripper, one end of the robot arm is disposed on the workbench, and the liquid transferring pump and the gripper are disposed at the other end of the robot arm.
Optionally, the system further comprises a refrigeration module arranged on the workbench and used for storing the extracted DNA product.
Optionally, the apparatus further comprises a shallow well plate and a micro-well plate, wherein the shallow well plate and the micro-well plate are both arranged on the plate position.
Optionally, the device further comprises a Tip head storage module arranged on the workbench and used for storing Tip heads required for transferring liquid.
Optionally, the device further comprises a constant temperature heating module arranged on the workbench and used for heating the lysis plate or the combination plate.
Optionally, the device further comprises an oscillation module, disposed on the working platform, for oscillating the sample in the lysis plate or the binding plate.
Optionally, the constant-temperature heating module and the oscillation module are of an integrated structure.
Optionally, the device further comprises a magnetic attraction device, which is arranged on the workbench and used for attracting magnetic beads used in DNA extraction.
A method of nucleic acid extraction using a dual extraction mode nucleic acid extraction station as described above, comprising the steps of:
s1, adding the lysis solution in the solution storage tank into the lysis plate with the sample stored therein by the mechanical arm device;
s2, the arm device moving the lysis plate to a centrifuge device to separate sample residue from lysis product;
s3, the mechanical arm device moves the cleavage products in the cleavage plate to a binding plate containing a binding solution and magnetic beads, and DNA is bound to the magnetic beads;
s4, transferring the combination board to a magnetic bar device by the mechanical arm device;
s5, adsorbing the magnetic beads combined with the DNA by the magnetic bar device;
s6, the mechanical arm device puts the combined board back to the board position;
s7, the mechanical arm device clamps a cleaning plate with cleaning solution stored therein, and the cleaning plate is placed on the magnetic bar device to clean the magnetic beads;
s8, the magnetic bar device adsorbs the magnetic beads, and the mechanical arm device clamps the cleaning plate and puts the cleaning plate back to the plate position;
s9, clamping an elution plate containing eluent by the mechanical arm device, placing the elution plate on the magnetic bar device, and eluting DNA combined in the magnetic beads;
and S10, adsorbing the magnetic beads by the magnetic bar device, clamping the elution plate by the mechanical arm device, and putting the elution plate back to the plate position to finish the DNA extraction work.
The invention has the beneficial effects that:
the invention provides a double-extraction-mode nucleic acid extraction workstation.A liquid storage tank and a plate position are arranged on a workbench, reagents required for DNA extraction are placed in the liquid storage tank, a cracking plate, a combination plate, an elution plate and a cleaning plate are placed on the plate position, a mechanical arm device adds the reagents in the liquid storage tank according to the DNA extraction process, the cracking plate can be moved to a centrifugal device to separate different products, then liquid is transferred to the combination plate, the combination plate is transferred to a magnetic rod device, and finally, the liquid elution plate containing DNA is transferred to the plate position to complete the DNA extraction. Through the arrangement, the automatic operation of DNA extraction can be realized, only a sample needs to be added into the lysis plate, and the difficulty of DNA extraction is reduced.
The invention also provides a nucleic acid extraction method, which utilizes the double-extraction-mode nucleic acid extraction workstation to realize automatic operation of DNA extraction and reduce the difficulty of DNA extraction.
Drawings
FIG. 1 is a schematic diagram of a dual extraction mode nucleic acid extraction workstation according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a stage in a dual extraction mode nucleic acid extraction station according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a robot device in a nucleic acid extraction workstation with dual extraction modes according to an embodiment of the present invention.
In the figure:
1-a robotic arm device; 11-clamping the hand; 2-a workbench; 21-plate position; 22-a liquid storage tank; 3-a centrifugal device; 4-a refrigeration module; 5-a vibration module; 6-Tip head storage module; 7-square hole plate; 8-magnetic bar device.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings and the embodiment. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention. It should be further noted that, for the convenience of description, only some but not all of the elements associated with the present invention are shown in the drawings.
In the description of the present invention, it should be noted that unless otherwise explicitly stated or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection or a removable connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In order to realize the automatic operation of DNA extraction, reduce the difficulty of DNA extraction and effectively ensure the smooth extraction of DNA, the invention provides a double-extraction-mode nucleic acid extraction workstation as shown in figures 1-3. The double extraction mode nucleic acid extraction station comprises two modes of operation, namely a pipetting mode and a magnetic rod mode.
Example one
This two mode nucleic acid extraction workstations of drawing adopts the bar magnet mode to carry out DNA extraction work, includes: the device comprises a workbench 2, a centrifugal device 3, a magnetic rod device 8 and a mechanical arm device 1, wherein the workbench 2 is provided with a liquid storage tank 22 and a plate position 21, the liquid storage tank 22 is used for storing reagents required by DNA extraction, and in the embodiment, the reagents mainly stored in the liquid storage tank 22 comprise lysis solution, binding solution, washing solution and eluent; the plate position 21 is used for placing a cracking plate, a combination plate, a cleaning plate and an elution plate; in order to store the waste liquid, a square hole plate 7 for storing the waste liquid is arranged on the plate position 21. If the DNA is required to be detected subsequently, a micro-porous plate and a shallow pore plate are placed on the plate position 21; the centrifugal device 3 is used for separating different products in the sample which is well cracked in the cracking plate under the centrifugal action; the magnetic bar device 8 is arranged on the workbench 2 and is used for adsorbing magnetic beads used in DNA extraction; the arm device 1 is disposed on the table 2, and can add a reagent in the solution storage tank 22 to the lysis plate, and can displace the lysis plate, the binding plate, the cleaning plate, or the elution plate.
Further, the mechanical arm device 1 comprises a mechanical arm, a liquid transferring pump and a clamping hand 11, wherein one end of the mechanical arm is arranged on the workbench 2, and the liquid transferring pump and the clamping hand 11 are arranged at the other end of the mechanical arm. The arm can shift combination board, elution board or lysis board through driving tong 11, and the liquid-transfering pump can be with being arranged in the reagent of depositing cistern 22 and adding lysis board or combination board, mutually supports through liquid-transfering pump and tong 11, guarantees that DNA draws and goes on smoothly. Moreover, the gripper 11 and the liquid-transferring pump are integrated on one mechanical arm, so that the utilization rate of the mechanical arm is improved. Of course, in other embodiments, the pipetting pump may be disposed on one mechanical arm, the gripper 11 may be disposed on the other mechanical arm, and the two mechanical arms cooperate to complete the DNA extraction operation, which is not limited herein.
Further, the double-extraction-mode nucleic acid extraction workstation further comprises a refrigeration module 4, wherein the refrigeration module 4 is arranged on the workbench 2 and used for storing the extracted DNA product. Effectively ensures that the DNA product can be preserved for a long time in a low-temperature environment.
Further, in order to prevent the occurrence of contamination during the transfer of the liquid, the double extraction mode nucleic acid extraction station further comprises a Tip head storage module 6 disposed on the table 2 for storing the Tip heads required for the transfer of the liquid. In this embodiment, the disposable Tip is replaced every time a pipetting operation is performed.
Further, the double-extraction-mode nucleic acid extraction workstation further comprises a constant-temperature heating module which is arranged on the workbench 2 and used for heating the lysis plate or the combination plate. After a sample is added into the cracking plate, a liquid transfer pump adds a cracking liquid into the cracking plate, the cracking plate is placed into the first constant-temperature heating module through the clamping hand 11, and the cracking speed of the sample can be accelerated through heating; the combination plate is transferred to a constant-temperature heating module for heating, so that the combination speed of the magnetic beads and the DNA can be increased.
Further, this two extraction mode nucleic acid extraction workstations still include and shake module 5, set up on workstation 2 for vibrate the sample in the lysis board or the combination board. The sample decomposition can be accelerated by oscillating the lysis plate, and the binding speed of DNA and magnetic beads can be accelerated by oscillating the binding plate. In this embodiment, optionally, the constant temperature heating module and the oscillation module 5 are of an integrated structure. Through the arrangement, the lysis plate or the combination plate can be vibrated and heated at the same time, so that the extraction speed of DNA is increased; and the occupied space can be reduced, so that the whole double-extraction-mode nucleic acid extraction workstation is more compact.
The embodiment also provides a nucleic acid extraction method, which uses the double-extraction-mode nucleic acid extraction workstation, and comprises the following steps:
s1, the mechanical arm drives the liquid transfer pump to automatically load and unload the Tip head, and the lysis solution in the solution storage tank 22 is added into the lysis plate;
s2, moving the lysis plate to the centrifugal device 3 by the clamping hand 11 to separate the sample residue from the lysis product; in order to accelerate the cracking speed, before centrifugation, the cracking plate can be transferred to a constant-temperature heating module and a vibration module 5 for heating and vibration;
s3, placing the centrifuged lysis plate on the plate position 21 by the gripper 11, discarding the upper plate of the discarded lysis plate in a garbage can to obtain a lysis product, and moving the lysis product in the lysis plate to a binding plate storing binding liquid and magnetic beads by a liquid transfer pump to combine DNA with the magnetic beads;
s4, transferring the combination board to the magnetic bar device 8 by the clamping hand 11;
s5, adsorbing the magnetic beads combined with the DNA by the magnetic bar device 8;
s6, the clamping hand 11 puts the combined board back to the board position 21;
s7, the cleaning plate with the cleaning solution is clamped by the clamp 11 and placed on the magnetic bar device 8, and the magnetic beads are cleaned;
s8, after the cleaning is finished, the magnetic bar device 8 adsorbs magnetic beads, and the mechanical arm device 1 clamps the cleaning plate and puts the cleaning plate back to the plate position 21;
the above steps S7 and S8 are repeated two or three times, and it is noted that a new cleaning plate needs to be replaced in each cleaning process;
s9, the gripper 11 grips the elution plate with the eluent and places the elution plate on the magnetic bar device 8, and DNA combined in the magnetic beads is eluted;
s10, the magnetic bar device 8 adsorbs magnetic beads, the gripper 11 grips the elution plate and puts the elution plate back to the plate position 21, and the DNA extraction work is completed.
Example two
The present embodiment provides a dual extraction mode nucleic acid extraction workstation, which works in a pipetting mode, the greatest difference between the pipetting mode and the magnetic bar mode is that a pipetting pump is needed to add lysis solution, binding solution, washing solution and eluent needed in the DNA extraction process according to the extraction steps, and the dual extraction mode nucleic acid extraction workstation further comprises a magnetic attraction device arranged on the workstation 2 for adsorbing magnetic beads used in DNA extraction.
The DNA extraction operation is carried out in a pipetting mode using the double extraction mode nucleic acid extraction workstation as described above, comprising the steps of:
s1, adding the lysis solution in the solution storage tank 22 into a lysis plate containing a sample by a liquid transfer pump;
s2, moving the lysis plate to the centrifugal device 3 by the clamping hand 11 to separate the sample residue from the lysis product;
s3, transferring the liquid in the lysis plate to the combination plate by a liquid transfer pump;
s4, adding a binding solution and magnetic beads into the binding plate to bind the DNA with the magnetic beads;
s5, transferring the combined plate to a magnetic suction device by the clamping hand 11;
s6, removing the waste binding liquid in the binding plate by the mechanical arm device 1, and adding the washing liquid into the binding plate;
s7, sucking out liquid obtained after washing the magnetic beads in the binding plate by a liquid transfer pump;
s8, adding the eluent into the binding plate by a liquid-transfer pump, so that the DNA is separated from the magnetic beads;
s9, sucking out the liquid containing the DNA by a liquid transfer pump, transferring the liquid to a DNA extraction plate, and finishing the DNA extraction work.
According to the double-extraction-mode nucleic acid extraction workstation provided by the invention, the liquid storage tank 22 and the plate position 21 are arranged on the workbench 2, reagents required for DNA extraction are placed in the liquid storage tank 22, the lysis plate, the combination plate, the elution plate and the cleaning plate are placed on the plate position 21, the mechanical arm device 1 is used for adding the reagents in the liquid storage tank 22 according to a DNA extraction process, the lysis plate can be moved to the centrifugal device 3 to separate different products, then the liquid is transferred to the combination plate, the combination plate is transferred to the magnetic rod device 8, and the liquid elution plate finally containing DNA is transferred to the plate position 21 to complete the DNA extraction. Through the arrangement, the automatic operation of DNA extraction can be realized, only a sample needs to be added into the lysis plate, and the difficulty of DNA extraction is reduced.
It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (10)
1. A dual extraction mode nucleic acid extraction workstation, comprising:
the device comprises a workbench (2), wherein a liquid storage tank (22) and a plate position (21) are arranged on the workbench (2), the liquid storage tank (22) is used for placing reagents required by DNA extraction, and the plate position (21) is used for placing a lysis plate, a combination plate, a cleaning plate and an elution plate;
a centrifugal device (3) for separating different products in the sample lysed in the lysis plate under the centrifugal action;
the magnetic bar device (8) is arranged on the workbench (2) and is used for adsorbing magnetic beads used in DNA extraction;
and the mechanical arm device (1) is arranged on the workbench (2), can add the reagent in the liquid storage tank (22) into the cracking plate, and can shift the cracking plate, the combination plate or the cleaning plate.
2. The dual extraction mode nucleic acid extraction station according to claim 1, wherein the robot arm device (1) comprises a robot arm, a pipetting pump, and a gripper (11), one end of the robot arm being disposed on the stage (2), and the pipetting pump and the gripper (11) being disposed at the other end of the robot arm.
3. The dual extraction mode nucleic acid extraction station of claim 1, further comprising a refrigeration module (4) disposed on the stage (2) for storing extracted DNA products.
4. The dual extraction mode nucleic acid extraction workstation of claim 1, further comprising a shallow well plate and a microplate, both of which are disposed on the plate location (21).
5. The dual extraction mode nucleic acid extraction station of claim 1, further comprising a Tip head storage module (6) disposed on the stage (2) for storing Tip heads required for transferring liquid.
6. The dual extraction mode nucleic acid extraction station of claim 1, further comprising a constant temperature heating module disposed on the stage (2) for heating the lysis plate or the binding plate.
7. The dual extraction mode nucleic acid extraction station of claim 6, further comprising a shaking module (5) disposed on the stage (2) for shaking the sample in the lysis plate or the binding plate.
8. The dual extraction mode nucleic acid extraction workstation of claim 7, wherein the constant temperature heating module is of unitary construction with the oscillation module (5).
9. The dual extraction mode nucleic acid extraction station of claim 1, further comprising a magnetic attraction device disposed on the stage (2) for attracting magnetic beads used in DNA extraction.
10. A method for nucleic acid extraction using the dual extraction mode nucleic acid extraction station of any one of claims 1-9, comprising the steps of:
s1, adding the lysis solution in the solution storage tank (22) into the lysis plate with the sample by the mechanical arm device (1);
s2, the arm device (1) moving the lysis plate to a centrifuge device (3) separating the sample residue from the lysis product;
s3, the mechanical arm device (1) moves the cleavage products in the cleavage plate to a binding plate containing a binding solution and magnetic beads, and DNA is bound to the magnetic beads;
s4, the mechanical arm device (1) transfers the combination board to a magnetic bar device (8);
s5, adsorbing the magnetic beads with DNA bonded by the magnetic bar device (8);
s6, the mechanical arm device (1) puts the combined board back to the board position (21);
s7, the mechanical arm device (1) clamps a cleaning plate with cleaning solution stored therein, places the cleaning plate on the magnetic bar device (8), and cleans the magnetic beads;
s8, adsorbing the magnetic beads by the magnetic bar device (8), and clamping the cleaning plate by the mechanical arm device (1) and putting the cleaning plate back to the plate position (21);
s9, the mechanical arm device (1) clamps an elution plate stored with eluent and places the elution plate on the magnetic bar device (8) to elute the DNA combined in the magnetic beads;
s10, adsorbing the magnetic beads by the magnetic bar device (8), clamping the elution plate by the mechanical arm device (1), and putting the elution plate back to the plate position (21) to finish the DNA extraction work.
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Application publication date: 20200724 |