Library construction workstation

Abstract

The utility model provides a library construction workstation, comprising: a nucleic acid extraction module for extracting nucleic acid; the quality inspection module is used for carrying out quality inspection on the extracted nucleic acid; the library building module and the quality inspection module are arranged side by side, and the library building module is used for carrying out library building on the nucleic acid subjected to quality inspection; the first transfer mechanism is used for transferring the nucleic acid extracted by the nucleic acid extraction module to the quality inspection module; and the second transfer mechanism is used for transferring the nucleic acid in the quality inspection module to the library building module. According to the technical scheme, through the cooperation of the nucleic acid extraction module, the quality inspection module, the library building module and the first transfer mechanism, the operations of nucleic acid extraction, quality inspection, library building, quality inspection, homogenization and the like are realized, the automation degree of library building is improved, the consistency of gene sequencing is further improved, and the influence of external risks on a detection result is reduced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of gene detection, and particularly relates to a library construction workstation. BACKGROUND

[0002] Gene sequencing technology is the most commonly used technology in modern molecular biology research, but a complex processing procedure needs to be performed between sample extraction and machine sequencing, including nucleic acid extraction, quality detection, library construction, quality detection, and uniformization operations. There are two traditional methods for the procedure: one is manual operation, which has the disadvantages of high labor intensity, easy pollution, and poor consistency; and the second is to use a traditional nucleic acid extractor plus a single-function library construction workstation, which is semi-automatic operation, and the quality detection of the extracted product and the uniformization of the library construction product need to be completed manually, and the user experience is poor. CONTENT OF THE UTILITY MODEL

[0003] The present application provides a library construction workstation, which improves the automation effect of library construction.

[0004] The present application provides a library construction workstation, which comprises:

[0005] a nucleic acid extraction module for extracting nucleic acid;

[0006] a quality detection module for detecting the extracted nucleic acid;

[0007] a library construction module arranged side by side with the quality detection module and used for constructing a library from the detected nucleic acid;

[0008] a first transfer mechanism for transferring the nucleic acid extracted by the nucleic acid extraction module to the quality detection module;

[0009] a second transfer mechanism for transferring the nucleic acid in the quality detection module to the library construction module.

[0010] In the above technical solution, through the cooperation of the nucleic acid extraction module, the quality detection module, the library construction module, and the first transfer mechanism, the operations of nucleic acid extraction, quality detection, library construction, quality detection, and uniformization are realized, the automation degree of library construction is improved, and the consistency of the library is improved, thereby reducing the influence of external risks on the detection result.

[0011] In one specific implementation scheme, the quality detection module comprises a first tip box, a quality detection tube rack, a quality detection plate site, a quality detection reagent box, and a fluorescence quantifier; wherein,

[0012] the first tip box, the quality detection reagent box, and the quality detection plate site are arranged in a single row;

[0013] the fluorescence quantifier and the quality detection tube rack are arranged in a single row;

[0014] The first transfer mechanism is arranged between the quality inspection reagent box and the quality inspection plate site.

[0015] In a specific embodiment, the first transfer mechanism comprises a reciprocating device, a single-channel pipettor, and a rotating gripper; wherein,

[0016] The reciprocating device comprises an annular transmission belt arranged on one side of the quality inspection reagent box and the quality inspection tube rack, and two hole positions fixedly arranged on the annular transmission belt;

[0017] The rotating gripper is used to transfer the quality inspection tube between the quality inspection tube rack, the reciprocating device, and the fluorescence quantitative instrument, and can be used to open or close the quality inspection tube;

[0018] The single-channel pipettor is used to suck the pipette from the first pipette box and pipette on the extraction reagent box of the nucleic acid extraction module, the reciprocating device, the quality inspection reagent box, and the quality inspection plate site.

[0019] In a specific embodiment, the library construction module comprises a second pipette box, a shaking module, a reagent box, a sample loading plate site, a magnetic rack, and a refrigeration module; wherein,

[0020] The second pipette box, the reagent box, the refrigeration module, the magnetic rack, the sample loading plate site, and the shaking module are arranged in an array;

[0021] The second pipette box is arranged in multiple rows and arranged in a stepped manner; wherein the second pipette box forms a stepped structure to avoid the nucleic acid extraction module and the quality inspection module;

[0022] The sample loading plate site, the reagent box, and the shaking module are arranged in the same column, the magnetic rack is arranged on one side of the reagent box, and the reagent box, the magnetic rack, and the shaking module are used for magnetic bead purification in the library construction process;

[0023] The second transfer mechanism is used to transfer consumables carried by the shaking module between the quality inspection plate site, the sample loading plate site, the magnetic rack, and the shaking module; wherein,

[0024] The consumables at least include a reaction plate and a purification plate.

[0025] In a specific embodiment, the second transfer mechanism comprises a three-axis motion platform, a rotating plate gripper arranged on the three-axis motion platform, and a multi-channel pipettor; wherein,

[0026] The multi-channel pipette is used to suck the tips in the second tip box and load reagents in the reagent tubes carried by the reagent box, the refrigeration module, the magnetic stand, the oscillation module and the sample plate site.

[0027] In a specific implementation, an information acquisition module is further arranged on the multi-channel pipette, and the information acquisition module is used to acquire information of the reagent box when the pipette collects reagents.

[0028] In a specific implementation, a rack is further arranged, and a workbench is arranged on the rack, and the nucleic acid extraction module, the quality inspection module and the library construction module are arranged on the workbench.

[0029] In a specific implementation, an air filtering device is further arranged on the top of the rack.

[0030] In a specific implementation, a thermal cycle module is further arranged, and the thermal cycle module is fixed on the workbench and used to provide temperature control for the library construction module.

[0031] In a specific implementation, a waste box is further arranged, and the waste box is arranged below the workbench, and an opening of the waste box is arranged diagonally to the nucleic acid extraction module. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 A structural schematic diagram of a library construction workstation provided by an embodiment of the present application is shown;

[0033] Figure 2 A schematic diagram of arrangement of modules of a library construction workstation provided by an embodiment of the present application is shown;

[0034] Figure 3 A layout schematic diagram of a quality inspection module provided by an embodiment of the present application is shown;

[0035] Figure 4 A side view of a library construction workstation provided by an embodiment of the present application is shown;

[0036] Figure 5 Another angle side view of a library construction workstation provided by an embodiment of the present application is shown. DETAILED DESCRIPTION

[0037] In order to make the purpose, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to the drawings.

[0038] Unless otherwise defined, technical terms and scientific terms used in one or more embodiments of the present disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Unless specifically mentioned otherwise, the use of the "first", "second" and similar words in one or more embodiments of the present disclosure does not indicate any order, quantity or importance, but is only used to distinguish different components. "Include" or "contain" and similar words mean that the elements or objects before the word cover the elements or objects listed after the word and their equivalents, and do not exclude other elements or objects. "Connected" or "connected" and similar words are not limited to physical or mechanical connections, but can include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to represent relative positional relationships, and when the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0039] To facilitate the understanding of the library construction workstation provided by the embodiments of the present application, the application scenario thereof is first described. The library construction workstation provided by the embodiments of the present application is used for gene sequencing. In the current detection mode, the process from sample extraction to sequencing is relatively complex and mainly relies on manual operation, resulting in poor consistency of detection results. Therefore, the embodiments of the present application provide a library construction workstation to improve the automation during library construction. The following will be described in detail in combination with specific drawings and embodiments.

[0040] Reference is made to Figure 1 , Figure 2 and Figure 4 and Figure 5 , Figure 1 shows a structural schematic diagram of the library construction workstation provided by the embodiments of the present application; Figure 2 shows a schematic diagram of the arrangement of each module of the library construction workstation provided by the embodiments of the present application; Figure 4 and Figure 5Different angle side views of the library construction workstation provided by the embodiments of the present application are shown. The library construction workstation provided by the embodiments of the present application mainly comprises a nucleic acid extraction module 10, a quality inspection module 20, a first transfer mechanism 30, a second transfer mechanism 50 and a library construction module 40. The nucleic acid extraction module 10 is used to extract nucleic acid, the quality inspection module 20 is used to inspect the nucleic acid, and the library construction module 40 is used to construct a library for the nucleic acid after the quality inspection. When the nucleic acid is transferred, the first transfer mechanism 30 is used to transfer the nucleic acid after extraction or the quality inspection tube after adding nucleic acid in each module, so as to realize the transfer of the nucleic acid between each module. Specifically, when the nucleic acid is transferred, the first transfer mechanism 30 is used to transfer the nucleic acid (also referred to as extraction product) extracted by the nucleic acid extraction module 10 to the quality inspection module 20. In addition, when the nucleic acid is inspected by the quality inspection module 20, the second transfer mechanism 50 is used to transfer the nucleic acid after the quality inspection to the library construction module 40. Thus, the nucleic acid extraction, quality inspection and library construction are realized.

[0041] As can be seen from the above description, the library construction workstation provided by the embodiments of the present application realizes the nucleic acid extraction, quality inspection, library construction, quality inspection and homogenization operation by the cooperation of the nucleic acid extraction module, the quality inspection module, the library construction module and the first transfer mechanism and the second transfer mechanism, improves the automation degree of the library construction, and further improves the consistency of the library and reduces the influence of external risks on the detection result.

[0042] The structure schematic diagram of the library construction workstation provided by the embodiments of the present application is described in detail below with reference to specific drawings.

[0043] Continuing to refer to the library construction workstation shown in Figure 1 and Figure 2 For the convenience of describing the arrangement between each module (the nucleic acid extraction module 10, the quality inspection module 20 and the library construction module 40), an XYZ coordinate system is constructed, wherein the X direction and the Y direction and the Z direction are perpendicular to each other, and the X direction and the Y direction are parallel to the placement plane of each module.

[0044] For the convenience of the first transfer mechanism 30 transferring the nucleic acid, when the nucleic acid extraction module 10, the quality inspection module 20 and the library construction module 40 are arranged, the first transfer mechanism 30 is located between the nucleic acid extraction module 10, the quality inspection module 20 and the library construction module 40. For example, when the arrangement is specifically arranged, the quality inspection module 20 and the library construction module 40 are arranged side by side (arranged along the X direction), and the nucleic acid extraction module 10 is located above the quality inspection module 20 (along the Y direction). When the first transfer mechanism 30 is arranged, the first transfer mechanism 30 is located between the quality inspection module 20 and the library construction module 40, and is opposite to the nucleic acid extraction module 10, that is, the quality inspection module 20, the library construction module 40 and the nucleic acid extraction module 10 are arranged around the first transfer mechanism 30, so that the first transfer mechanism 30 can transfer the nucleic acid between each module with a smaller path.

[0045] With reference to Figure 2 The nucleic acid extraction module 10 provided by the embodiments of the present application is used to extract nucleic acid, and can be a multi-channel magnetic rod nucleic acid extraction module 10 which can simultaneously perform high-quality nucleic acid extraction on multiple samples. Specifically, the multi-channel magnetic rod nucleic acid extraction module 10 can be a magnetic rod nucleic acid extraction instrument. The nucleic acid extraction module 10 has an extraction kit 11 which is used to carry various reagents required for nucleic acid extraction. In use, the product (containing nucleic acid) extracted by the nucleic acid extraction module 10 is carried by a test tube on the extraction kit 11. When the nucleic acid is transferred by the first transfer mechanism 30, the product can be sucked from the extraction kit 11 to suck the nucleic acid.

[0046] With reference to Figure 2 and Figure 3 The quality inspection module 20 provided by the embodiments of the present application mainly includes a first suction head box 21, a quality inspection reagent kit 22, a quality inspection tube rack 23, a fluorescent quantification instrument 24 and a quality inspection plate site 25. The first suction head box 21 is used to carry suction heads which are used in cooperation with the first transfer mechanism 30 to suck nucleic acid or reagents through the suction heads. The quality inspection tube rack 23 is used to carry quality inspection tubes (which are test tubes or other tubular structures capable of carrying nucleic acid) which are used to carry nucleic acid and reagents. The quality inspection reagent kit 22 is used to carry quality inspection and homogenization reagents. The fluorescent quantification instrument 24 is used to inspect the nucleic acid. The quality inspection plate site 25 is used as a transfer structure to transfer the nucleic acid after quality inspection to consumables "reaction plates".

[0047] In the specific arrangement of the above-mentioned first suction head box 21, quality inspection tube rack 23, quality inspection reagent kit 22, quality inspection plate site 25 and fluorescent quantification instrument 24, the first suction head box 21, quality inspection reagent kit 22 and quality inspection plate site 25 are arranged in a single row, specifically along the X direction. In the specific arrangement, the first suction head box 21 is located at the leftmost side, the quality inspection plate site 25 is located at the rightmost side, and the quality inspection reagent kit 22 is located between the two. The fluorescent quantification instrument 24 and the quality inspection tube rack 23 are arranged in a single row, specifically along the X direction. In the specific arrangement, the fluorescent quantification instrument 24 is located at the left side, and the quality inspection tube rack is located at the right side. Moreover, the fluorescent quantification instrument 24 and the quality inspection tube rack 23 are located below the first suction head box 21. In the specific arrangement, the nucleic acid extraction module 10 is located above the quality inspection reagent kit 22 and the first suction head box 21, and the first transfer mechanism 30 is arranged between the quality inspection reagent kit 22 and the quality inspection plate site 25. In the above-mentioned arrangement, the first transfer mechanism 30 can suck the suction heads in the first suction head box 21 and the product in the extraction kit 11 in the nucleic acid extraction module 10 through a shorter path. At the same time, the quality inspection reagents and homogenization reagents (such as ultrapure water) in the quality inspection reagent kit 22 can also be sucked through a smaller path.

[0048] With reference to Figure 3 andFigure 4 The first transfer mechanism 30 comprises a reciprocating device 35, a single-channel pipette 33 and a rotating gripper 34. The reciprocating device 35 is used to carry the quality inspection tube and comprises an annular conveying belt 352 and two hole positions 351 fixed on the annular conveying belt 352. The quality inspection tube can be placed on the hole position 351 when the quality inspection tube is carried. The annular conveying belt 352 is arranged in the length direction along the Y direction, and the annular plane formed thereby is parallel to the placement plane of the nucleic acid extraction module 10 and the quality inspection module 20. The annular conveying belt 352 is located between the quality inspection reagent box 22 and the quality inspection plate position 25, and when the annular conveying belt 352 rotates, the two hole positions 351 can be moved between the quality inspection reagent box 22 and the quality inspection tube rack 23. That is, the nucleic acid extraction module 10, the quality inspection reagent box 22, the quality inspection tube rack 24 and the quality inspection plate position 25 are all located outside the annular conveying belt 352, so that the quality inspection tube can be transplanted through a smaller moving path.

[0049] When the quality inspection tube is moved, the rotating gripper 34 is used to grab the quality inspection tube, and the rotating gripper 34 is used to transfer the quality inspection tube between the quality inspection tube rack 23, the reciprocating device 35 and the fluorescence quantitative instrument 24, and can be used to open or close the quality inspection tube. Specifically, the rotating gripper 34 is used to transplant the quality inspection tube on the quality inspection tube rack 23 to one hole position 351 on the annular conveying belt 352, so that the quality inspection tube can be placed on the hole position 351. In addition, the rotating gripper 34 is also used to transplant the quality inspection tube loaded with reagent to the fluorescence quantitative instrument 24, and transplant the quality inspection tube after completing the fluorescence quantitative detection to the quality inspection tube rack 23. So as to realize the transfer of the quality inspection tube between the reciprocating device 35, the fluorescence quantitative instrument 24 and the quality inspection tube rack 23.

[0050] When the nucleic acid or the uniformization reagent is sucked, the quality inspection tube can be opened by the rotating gripper 34, and then the single-channel pipette 33 is used to realize. The single-channel pipette 33 can be used to suck the suction head from the first suction head box 21, and pipette in the extraction reagent box 11 of the nucleic acid extraction module 10, the reciprocating device 35, the quality inspection reagent box 22 and the quality inspection plate position 25. In the implementation, the single-channel pipette 33 sucks the suction head from the first suction head box 21, and sucks the extraction product in the extraction reagent box 11 of the nucleic acid extraction module 10, and adds to the quality inspection tube carried on the hole position 351. In addition, it is also used to add the uniformization reagent in the quality inspection reagent box 22 to the quality inspection tube carried on the hole position 351. Then the quality inspection tube is closed by the rotating gripper 34.

[0051] It should be understood that in a specific implementation, the rotating gripper 34 and the single-channel pipette 33 are carried on two different three-axis motion platforms respectively, for the purpose of description, the three-axis motion platform carrying the single-channel pipette 33 is named as the first three-axis motion platform 31, and the three-axis motion platform carrying the rotating gripper 34 is named as the second three-axis motion platform 32. The first three-axis motion platform 31 and the second three-axis motion platform 32 can move in XYZ directions, wherein the Z direction is perpendicular to the X direction and the Y direction. When the rotating gripper 34 and the single-channel pipette 33 are in use, they can be moved to corresponding operation positions by the first three-axis motion platform 31 and the second three-axis motion platform 32. For example, the first three-axis motion platform 31 and the second three-axis motion platform 32 can include rails parallel to the X direction, the Y direction and the Z direction respectively, and the rails are slidably connected to each other.

[0052] When the extraction product is transferred to the quality inspection module for inspection by the first transfer mechanism, the specific process is as follows: adding a quality inspection reagent to a quality inspection tube, adding the extraction product to the quality inspection tube and mixing by blowing, detecting the mixture in the quality inspection tube to obtain a quality inspection result, and then adding the extraction product and a homogenization reagent to the reaction plate according to the quality inspection result and the library building and sample loading parameters.

[0053] When the quality inspection reagent is added to the quality inspection tube, the rotating gripper 34 of the first transfer mechanism 30 transfers the first quality inspection tube from the quality inspection tube rack 23 to the hole position of the reciprocating motion device 35, opens the tube cap, and puts the tube cap back into the hole position of the quality inspection tube rack 23;

[0054] The first quality inspection tube is moved to the working position of the single-channel pipette 33 by the reciprocating motion device 35, the single-channel pipette 33 loads a suction head in the first suction head box and sucks the quality inspection reagent from the quality inspection reagent box 22 into the first quality inspection tube, and then the first transfer mechanism 30 puts the first quality inspection tube back into the quality inspection tube rack 23, so that the reciprocating motion device 35 is used to add the quality inspection reagent to the second quality inspection tube, the third quality inspection tube and more quality inspection tubes in turn, and the first transfer mechanism 30 is used to put the second quality inspection tube, the third quality inspection tube and more quality inspection tubes back into the quality inspection tube rack 23. At the same time in this process, the nucleic acid extraction module 10 completes the extraction of nucleic acid to generate the extraction product, and the extraction product is located in the elution hole of the extraction reagent box 11.

[0055] Then when the extraction product is added to the quality inspection tube and mixed by blowing, the first quality inspection tube containing the quality inspection reagent is placed in the hole position of the reciprocating motion device 35 by the first transfer mechanism 30. The single-channel pipette 33 loads a suction head in the first suction head box and sucks the extraction product from the corresponding elution hole of the extraction reagent box 11, and adds it to the first quality inspection tube for mixing by blowing;

[0056] The second quality inspection tube is transferred from the quality inspection tube rack to another hole position of the reciprocating device 35 by the rotating gripper 34, and the tube cap is opened; the first quality inspection tube is moved to the position of the rotating gripper 34 by the reciprocating device 35, and the tube cap of the second quality inspection tube is covered on the first quality inspection tube; and the first quality inspection tube is moved to the hole position of the quality inspection tube rack by the rotating gripper 34;

[0057] After the first set time interval, the third quality inspection tube is grabbed from the quality inspection tube rack by the rotating gripper 34 and placed in the hole position of the reciprocating device 35; at the same time, the second quality inspection tube is loaded with extraction products and mixed by blowing by the single-channel pipettor 33;

[0058] After the first quality inspection tube is incubated for the second set time, the cabin door of the fluorescence quantitative instrument is opened, and the first quality inspection tube is placed into the fluorescence quantitative instrument by the rotating gripper 34 at the interval time, and after the detection is completed, the first quality inspection tube is taken out from the fluorescence quantitative instrument by the rotating gripper 34 and placed back to the corresponding position of the quality inspection tube rack;

[0059] The second quality inspection tube and the third quality inspection tube are sequentially sent into the fluorescence quantitative instrument for quality inspection, and after the quality inspection is completed, the second quality inspection tube and the third quality inspection tube are sent into the quality inspection tube rack.

[0060] As can be seen from the above description, the rotating gripper 34 and the single-channel pipettor 33 cooperate with the reciprocating device 35 to perform the operations of opening the cap, transferring, adding liquid and the like on multiple quality inspection tubes. For any quality inspection tube, the rotating gripper 34, the single-channel pipettor 33 and the reciprocating device 35 disclosed in the embodiment are not sent into the fluorescence quantitative instrument after completing the operations of adding quality inspection reagents, adding extraction products and mixing by blowing and being static, and then another quality inspection tube is operated, but another quality inspection tube is operated in the time interval of transferring, adding liquid or being static of the quality inspection tube, so as to improve the efficiency of the whole process.

[0061] For example, when the extraction product is detected, the rotating gripper 34 transfers the No. 1 detection tube from the detection tube rack 23 to the hole position 351 of the reciprocating device, and rotates the cover to open, and the rotating gripper 34 places the tube cover back into the No. 1 hole of the detection tube rack 23. At the same time, the reciprocating device 35 moves the detection tube to the working position of the single-channel pipettor 33 (the position of the detection reagent kit 22), and at the same time, the single-channel pipettor 33 loads a suction head in the first suction head box 21, sucks the detection reagent from the detection reagent kit 22 into the detection tube, and then the first transfer mechanism 30 places the detection tube back into the detection tube rack 23. In this way, the operation of adding detection reagent to multiple detection tubes is completed, and in this process, the nucleic acid extraction module 10 completes the extraction of nucleic acid to generate the extraction product, which is located in the extraction reagent kit 11. Then, the first transfer mechanism 30 places the detection tube containing the detection reagent into the hole position of the reciprocating device 35, the single-channel pipettor 33 loads a suction head in the first suction head box to suck the extraction product from the elution hole of the extraction reagent kit 11, and transfers it to the detection tube in the corresponding hole position 351 of the reciprocating device, and blows and mixes. At the same time, the rotating gripper 34 grabs the No. 2 detection tube from the detection tube rack 23, transfers it to the hole position 351 of the reciprocating device, and rotates the cover to open. Then the suction head is returned to the original position, and at the same time, the reciprocating device 35 transfers the detection tube to the operating position of the rotating gripper 34, which is screwed onto the No. 1 detection tube with the tube cover of the No. 2 detection tube by the rotating gripper 34, and the No. 1 detection tube is transferred to the No. 2 hole of the detection tube rack 23. After a certain period of time, the rotating gripper 34 grabs the No. 3 detection tube from the detection tube rack 23 and places it in the hole position 351 of the reciprocating device. During the time of closing the cover and grabbing a new tube, the single-channel pipettor 33 loads the sample for the No. 2 detection tube and blows and mixes, and the cycle is repeated. After the No. 1 detection tube is incubated for a proper period of time, the door of the fluorescence quantitative instrument 24 is opened, and the detection tube is grabbed by the rotating gripper 34 and placed in the fluorescence quantitative instrument 24, the door of the fluorescence quantitative instrument 24 is closed, and the detection instruction is sent by the upper computer, the fluorescence quantitative instrument 24 transmits the data to the upper computer for saving and processing after detection, and at the same time, the door of the fluorescence quantitative instrument 24 is opened, the detection tube is taken out from the fluorescence quantitative instrument 24 by the rotating gripper 34 and placed back into the corresponding position of the detection tube rack 23, and the cycle is repeated. In the above-mentioned scheme, the operation of multiple detection tubes is performed by using the transfer of each detection tube and the interval of liquid addition and static state, so that the first transfer mechanism 30 can reasonably utilize the time difference to increase the operation efficiency of the detection tube, and the detection efficiency of the fluorescence quantitative instrument 24 is improved.

[0062] After the extraction product quality inspection is completed, according to the quality inspection result and the library building sample loading parameter, a single channel pipettor 33 sucks a certain proportion of nucleic acid and uniformization reagent from the elution hole of the extraction reagent box 11 and the quality inspection reagent box 22, and adds them into the consumable “reaction plate” on the quality inspection plate site 25. The library building product quality inspection process is the same as above, only the sample position is changed from the extraction reagent box 11 to the quality inspection plate site 25; during the uniformization operation, according to the concentration quality inspection data in the upper computer, the single channel pipettor 33 sucks the uniformization reagent in the quality inspection reagent box 22, and adds it into the consumable “reaction plate” on the quality inspection plate site 25, so as to dilute the library building product to the required concentration for sequencing.

[0063] In an implementable scheme, the two hole sites 351 are arranged at intervals. The arrangement positions of the two hole sites 351 satisfy that when the quality inspection tube carried by one of the hole sites 351 is loaded with nucleic acid, the other hole site 351 can be moved to the working area of the rotary clamp jaw 34, so that the movement of the rotary clamp jaw 34 and the single channel pipettor 33 does not interfere with each other, so as to make full use of the time of loading sample and blowing and mixing by the single channel pipettor 33, and to carry out the operations of opening and closing the cap, transferring the quality inspection tube and the like by the rotary clamp jaw 34, thereby improving the detection efficiency.

[0064] Referring to Figure 2 and Figure 5 together, the library building module 40 provided in the embodiments of the present application is used to construct a library for nucleic acid. The library building module 40 provided in the embodiments of the present application mainly comprises a second suction head box 41, a shaking module 42, a reagent box 43, a sample loading plate site 45, a magnetic stand 46, a refrigeration module 48 and a second transfer mechanism 50. The second suction head box 41 is used to carry suction heads; the reagent box 43 is used to carry large volume reagents required for library building, the sample loading plate site 46 is used to maintain a low temperature environment when adding reagents into the consumable “reaction plate”, so as to prevent the reagents from reacting in advance, the refrigeration module 48 can refrigerate and store key reagents required for library building, so as to prevent deterioration; the magnetic stand 46 and the shaking module 42 are used for magnetic bead purification of nucleic acid, so as to meet the high efficiency magnetic bead purification in the library building process. Specifically, the reagent box 43, the magnetic stand 46 and the shaking module 42 are used for magnetic bead purification in the library building process. The second transfer mechanism 50 is used to flexibly transfer the consumables (which at least include a reaction plate and a purification plate) among the quality inspection plate site 25, the sample loading plate site 45, the thermal cycle module 47, the magnetic stand 46, the shaking module 42, and is used to load the reagents in the refrigeration module 48 and the reagent box 43 into the consumables “reaction plate” and “purification plate”. The functions of the above-mentioned second suction head box 41, reagent box 43, refrigeration module 48, magnetic stand 46 and shaking module 42 are all common modules in the library building module 40, which will not be described in detail in the embodiments of the present application.

[0065] In addition, the library building module 40 provided by the embodiments of the present application can further include a thermal cycling module 47 fixed on the workbench 100 and used to provide rapid and high-precision temperature control for the library building module 40. In a specific arrangement, the thermal cycling module 47 is also located at a corner of the workbench 100, and a blowing fan 301 is arranged at the right side of the thermal cycling module 47 to quickly extract the aerosol generated so as to reduce the risk of pollution.

[0066] In addition, the library building module 40 further includes a waste box 44 used to contain the waste generated in the library building process. In a specific arrangement, the waste box 44 is arranged diagonally with the nucleic acid extraction module 10 so as to reasonably utilize the space. In addition, the waste box 44 is located at a corner so as to be relatively far away from other modules, thereby reducing the probability of waste polluting other substances.

[0067] With reference to the arrangements of the various plate positions in the library building module 40 shown in Figure 2 , Figure 4 and Figure 5 , the second tip box 41, the shaking module 42, the reagent box 43, the waste box 44, the sample adding plate position 45, the magnetic stand 46, the thermal cycling module 47, and the refrigeration module 48 are arranged in an array. As shown in Figure 2 , the second tip box 41, the shaking module 42, the reagent box 43, the waste box 44, the sample adding plate position 45, the magnetic stand 46, the thermal cycling module 47, and the refrigeration module 48 are arranged in an array, and the whole forms a stepped structure to avoid the structures in the nucleic acid extraction module 10 and the quality inspection module 20, thereby making the various plate positions of the library building workstation form an arrayed whole structure on the placement surface.

[0068] In a specific arrangement, the second tip box 41 is arranged in multiple rows and in a stepped manner. The stepped second tip box 41 avoids the nucleic acid extraction module 10, the quality inspection module 20, and the refrigeration module 44. As shown in Figure 2 , the second tip box 41 is arranged in three rows, and each row is arranged along the Y direction. The multiple rows of second tip boxes 41 are arranged along the X direction. Along the X direction, the second tip box 41 in the first row is one, the second tip boxes 41 in the second row are five, and the second tip boxes 41 in the third row are three, thereby forming a stepped arrangement. When matched with the quality inspection module 20 and the nucleic acid extraction module 10, the second tip boxes 41 in the first row are arranged in a row along the Y direction with the quality inspection plate position 25. The second tip boxes 41 in the second row and the third row are located at one side of the nucleic acid extraction module 10, thereby reasonably utilizing the space and integrating the nucleic acid extraction module 10, the quality inspection module 20, and the library building module 40 on a placement surface, and making the three modules closely arranged.

[0069] The refrigeration module 48 has multiple, as shown in Figure 2As shown in the figure, the number of the refrigeration modules 42 is 4, and the 4 refrigeration modules 42 are arranged in two rows along the Y direction, and are located at the right side of the second row of the second tip box 41, above the third row of the second tip box 41 and the sample plate position 45.

[0070] The sample plate position 45, the reagent box 43 and the oscillation module 42 are arranged in a row along the Y direction, and in terms of sequence, from top to bottom, they are the sample plate position 45, the reagent box 43 and the oscillation module 42 respectively. The thermal cycle module 47 and the magnetic stand 46 are arranged in a row along the Y direction, and in terms of sequence, from top to bottom, they are the thermal cycle module 47 and the magnetic stand 46 respectively.

[0071] In the library building process, the consumables “reaction plate” and “purification plate” can be transferred between the quality inspection plate position 25, the sample plate position 45, the thermal cycle module 47, the magnetic stand 46 and the oscillation module 42 by the second transfer mechanism, and the reagents in the refrigeration module 48 and the reagent box 43 can be loaded into the consumables “reaction plate” and “purification plate”.

[0072] In a specific arrangement, the second transfer mechanism 50 includes a multi-channel pipette 52, which is used to suck the tips in the second tip box 41 and to transfer the liquid in the library building module.

[0073] In a specific use, the second transfer mechanism 50 includes a three-axis motion platform, which is named as the third three-axis motion platform 51 for convenience of description. The third three-axis motion platform 51 can move in the X direction, the Y direction and the Z direction. In addition, the second transfer mechanism 50 further includes a transfer plate clamping jaw 53 arranged on the third three-axis motion platform 51, and the multi-channel pipette 52 is also arranged on the third three-axis motion platform 51. The transfer plate clamping jaw 53 can flexibly transfer the consumables “reaction plate” and “purification plate” between the quality inspection plate position 25, the sample plate position 45, the thermal cycle module 47, the magnetic stand 46 and the oscillation module 42.

[0074] In an optional case, the library building module 40 further includes an information acquisition module 54 arranged on the multi-channel pipette 52, which is used to acquire the information of the reagent box when the pipette collects the reagent. The information is uploaded to the upper computer to realize the acquisition of the information. In addition, the information acquisition module 54 can also acquire the information of the consumables and the position information of the motion mechanism (the second transfer mechanism 50). The upper computer judges whether the consumables are misplaced or missed according to the information acquired by the information acquisition module 54, and calibrates the position deviation of the motion mechanism according to the information acquired by the information acquisition module 54. It should be understood that the execution of the upper computer according to the information acquired by the information acquisition module 54 is a conventional control mode in the art, which will not be described here.

[0075] To carry the above-mentioned various modules, the library construction workstation provided by the embodiment of the application further comprises a rack 200, and the rack 200 is provided with a workbench 100, and the nucleic acid extraction module 10, the quality inspection module 20 and the library construction module 40 are all arranged on the workbench 100. The above-mentioned modules are carried by the workbench 100. In addition, the three-axis motion platforms in the first transfer mechanism 30 and the second transfer mechanism 50 can be supported by the rack 200, and the transfer plate clamping jaw 53 and the rotary clamping jaw 33 are suspended above the workbench 100, so that the clamping and taking of the quality inspection tube can be realized to move between different modules.

[0076] The rack 200 is further provided with a sealed cabin door 201, and the sealed cabin door 201 and the rack instrument isolate the air in the instrument from the outside of the instrument, so as to prevent sample pollution caused by the external environment, and also prevent nucleic acid pollution in the instrument to the external environment.

[0077] In an optional scheme, the library construction workstation further comprises an air filtration device 300 arranged on the top of the rack 200. The air filtration device 300 is installed on the top of the rack 200, and has a fan and a high-efficiency filter, which can filter and purify the external air and then send it into the cabinet to provide a clean positive pressure environment in the cabinet, prevent aerosol pollution, and correspondingly, the left and right sides of the rack 200 are provided with air outlet fans 301, and the air volume of the air outlet fans 301 is less than that of the air filtration device 300.

[0078] In order to facilitate the understanding of the library construction workstation provided by the embodiment of the application, the embodiment of the application further provides a nucleic acid library construction method, which utilizes any one of the library construction workstations described above; the method comprises the following steps:

[0079] Step 001: performing nucleic acid extraction by the nucleic acid extraction module and forming an extraction product;

[0080] Specifically, reference is made to the related description in Figures 1-5 and details are not repeated here.

[0081] Step 002: transferring the extraction product to the quality inspection module by the first transfer mechanism for quality inspection;

[0082] Specifically, the following steps are included:

[0083] Step a, adding a quality inspection reagent to the quality inspection tube;

[0084] Specifically, the quality inspection tube is transferred from the quality inspection tube rack to the hole position of the reciprocating motion device by the rotary clamping jaw of the first transfer mechanism, and the tube cap is rotated open.

[0085] The quality inspection tube is moved to the working position of the single-channel pipette by the reciprocating motion device; the single-channel pipette loads the pipette tip in the first pipette tip box and sucks the quality inspection reagent from the quality inspection reagent box to add to the quality inspection tube; then the first transfer mechanism puts the quality inspection tube back to the quality inspection tube rack;

[0086] The above steps are repeated until the quality inspection reagent is added to the predetermined number of quality inspection tubes.

[0087] Step b, adding the extraction product to the quality inspection tube and mixing by blowing;

[0088] Specifically, the first quality inspection tube is placed into the hole position of the reciprocating motion device by the first transfer mechanism, the tube cap is rotated to open, and the tube cap is put back into the hole position of the quality inspection tube rack; the single-channel pipette loads the pipette tip in the first pipette tip box and sucks the extraction product from the corresponding elution hole of the extraction reagent box and adds to the first quality inspection tube to mix by blowing;

[0089] The second quality inspection tube is transferred from the quality inspection tube rack to another hole position of the reciprocating motion device by rotating the clamping jaw, and the tube cap is rotated to open; the first quality inspection tube is moved to the position of the clamping jaw by the reciprocating motion device, and the tube cap of the second quality inspection tube is covered on the first quality inspection tube; and the first quality inspection tube is moved to the hole position of the quality inspection tube rack by the clamping jaw;

[0090] After a first set time interval, the third quality inspection tube is grabbed from the quality inspection tube rack by the clamping jaw and placed in the hole position of the reciprocating motion device; at the same time, the single-channel pipette sucks the extraction product from the corresponding elution hole of the extraction reagent box and adds to the second quality inspection tube to mix by blowing;

[0091] The above steps are repeated until the extraction product of the predetermined number of quality inspection tubes is added to the corresponding elution hole and mixed by blowing.

[0092] Step c, detecting the quality inspection result of the mixture of the quality inspection tube;

[0093] Specifically, after the first quality inspection tube is incubated for a second set time, the cabin door of the fluorescence quantitative instrument is opened, the first quality inspection tube is taken out by the clamping jaw and put into the fluorescence quantitative instrument for detection at an interval, and after the detection is completed, the first quality inspection tube is taken out from the fluorescence quantitative instrument by the clamping jaw and put back to the corresponding position of the quality inspection tube rack;

[0094] The above steps are repeated until the quality inspection of the predetermined number of quality inspection tubes is completed.

[0095] Step d, adding the extraction product and the uniformization reagent to the reaction plate according to the quality inspection result and the library building and sample loading parameters.

[0096] According to the quality inspection results of each quality inspection tube and the library building sample parameters, the single-channel pipettor respectively sucks a certain proportion of extraction products and uniformization reagents from the elution holes of the extraction kit and the quality inspection kit, and adds them into the corresponding hole positions of the reaction plate on the quality inspection plate position.

[0097] Specific reference Figures 1-5 is made to the relevant description in the foregoing, which will not be repeated here.

[0098] Step 003: transferring the nucleic acid after quality inspection of the quality inspection module to the library building module for library building by the second transfer mechanism.

[0099] Specific reference Figures 1-5 is made to the relevant description in the foregoing, which will not be repeated here.

[0100] In the above technical solution, through the cooperation of the nucleic acid extraction module, the quality inspection module, the library building module and the first transfer mechanism, nucleic acid extraction, quality inspection, library building, quality inspection, uniformization and other operations are realized, the automation degree of library construction is improved, and the consistency of the library is improved, thereby reducing the influence of external risks on the detection results.

[0101] One or more embodiments of the present specification are intended to cover all such alternatives, modifications and variations falling within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent replacements, improvements and the like made in the spirit and principles of one or more embodiments of the present specification should be included in the protection scope of the present disclosure.

[0102] The above is merely a specific implementation of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can easily think of changes or replacements within the technical range disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A document library construction workstation, characterized in that, include: The nucleic acid extraction module is used to extract nucleic acids. The quality control module is used to perform quality control on the extracted nucleic acids; The library construction module is arranged side by side with the quality inspection module and is used to construct a library from the nucleic acids after quality inspection. The first transfer mechanism is used to transfer the nucleic acid extracted by the nucleic acid extraction module to the quality inspection module; The second transfer mechanism is used to transfer the nucleic acids in the quality inspection module to the library construction module.

2. The document construction workstation according to claim 1, characterized in that, The quality control module includes: a first pipette tip box, a quality control tube rack, a quality control plate position, a quality control reagent kit, and a fluorescence quantitative PCR instrument; wherein... The first suction tip box, the quality control reagent kit, and the quality control plate are arranged in a single row. The fluorescence quantitative instrument and the quality inspection tube rack are arranged in a single row; The first transfer mechanism is disposed between the quality control kit and the quality control plate position.

3. The document construction workstation according to claim 2, characterized in that, The first transfer mechanism includes: a reciprocating motion device, a single-channel pipette, and a rotating gripper; wherein, The reciprocating motion device includes an annular conveyor belt located on one side of the quality inspection kit and the quality inspection tube rack, and two holes fixed at intervals on the annular conveyor belt; The rotating gripper is used to transfer the quality inspection tube between the quality inspection tube rack, the reciprocating motion device and the fluorescence quantitative instrument, and can also be used to open or close the quality inspection tube; The single-channel pipette is used to draw pipette tips from the first pipette tip box and pipette them onto the extraction kit, the reciprocating motion device, the quality control kit, and the quality control plate of the nucleic acid extraction module.

4. The document construction workstation according to claim 3, characterized in that, The library construction module includes: a second pipette tip box, a shaking module, a reagent kit, a sample application slot, a magnetic rack, and a refrigeration module; wherein... The second pipette tip box, the reagent kit, the refrigeration module, the magnetic rack, the sample dispensing plate, and the shaking module are arranged in an array. The second pipette tip boxes are arranged in multiple rows in a stepped manner; wherein the stepped arrangement of the second pipette tip boxes avoids the nucleic acid extraction module and the quality inspection module. The sample loading plate, the reagent kit, and the shaking module are arranged in the same column, the magnetic rack is located on one side of the reagent kit, and the reagent kit, the magnetic rack, and the shaking module are used for magnetic bead purification during the library construction process; The second transfer mechanism is used to transfer consumables carried on the quality inspection plate and the vibration module between the quality inspection plate, the sample application plate, the magnetic rack, the refrigeration module, and the vibration module; wherein, The consumables include at least reaction plates and purification plates.

5. The document construction workstation according to claim 4, characterized in that, The second transfer mechanism includes a three-axis motion platform, and a rotating plate gripper and a multi-channel pipette disposed on the three-axis motion platform; wherein, The multichannel pipette is used to aspirate pipette tips in the second tip box and to load reagents into the reagent tubes held in the reagent kit, the refrigeration module, the magnetic rack, the shaker, and the sample loading plate.

6. The document construction workstation according to claim 5, characterized in that, It also includes an information acquisition module disposed on the multichannel pipette, the information acquisition module being used to acquire information about the reagent kit when the pipette collects reagents.

7. The library construction workstation according to any one of claims 1 to 6, characterized in that, It also includes a rack, on which a workbench is provided, and the nucleic acid extraction module, the quality inspection module and the library construction module are all located on the workbench.

8. The document construction workstation according to claim 7, characterized in that, It also includes an air filtration device, which is located on top of the frame.

9. The document construction workstation according to claim 8, characterized in that, It also includes a heat circulation module, which is fixed on the workbench and used to provide temperature control for the database construction module.

10. The library construction workstation according to claim 8, characterized in that, It also includes a waste box, which is located below the workbench; and the opening of the waste box is diagonally positioned with respect to the nucleic acid extraction module.

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