CN107916222B

CN107916222B - Accurate split charging and mixing device for DNA primers or probes and application method thereof

Info

Publication number: CN107916222B
Application number: CN201711270989.8A
Authority: CN
Inventors: 李俊
Original assignee: Briggs Biotechnology Shanghai Co ltd
Current assignee: Briggs Biotechnology Shanghai Co ltd
Priority date: 2017-12-05
Filing date: 2017-12-05
Publication date: 2023-10-10
Anticipated expiration: 2037-12-05
Also published as: CN107916222A

Abstract

The invention discloses a precise split charging and mixing device of a DNA primer or a probe and a use method thereof, wherein the precise split charging and mixing device mainly comprises a control cabinet, an actuating mechanism, a working area part, a testing unit, a gun head frame, a working platen and a fixed platen; the actuating mechanism consists of an X-axis sliding table, a Y-axis sliding table, a Z-axis sliding table and a gun-axis sliding table; the working area part comprises 8 split charging target plates, 1 gun head plate, 1 stock solution plate, 1 liquid guiding target plate and 1 gun head separator, and the working table plate adopts a layout form taking the stock solution plate as the center. The invention provides a precise, reliable and efficient engineering solution, thereby realizing automatic and precise split charging and mixing of DNA primers or probes. The invention can save labor, can save tedious manual liquid sucking and pumping processes, and an operator only needs to use the mobile USB flash disk to guide the required operation into software, so that the previous preparation work is saved, and the full-automatic operation is realized.

Description

Accurate split charging and mixing device for DNA primers or probes and application method thereof

Technical Field

The invention belongs to the technical field of molecular biology, and particularly relates to a precise split charging and mixing device for a DNA primer or a probe and a use method thereof.

Background

In molecular biology research, primers having different base sequences and lengths are required to be synthesized according to specific purposes. After synthesis, the DNA primers are typically in a standard well plate, such as a 48-well plate or a 96-well plate, and a different primer may be present in each well. Depending on the purpose of analysis and investigation, it may be necessary to dispense the same (called a single) synthesized primer or probe into several different centrifuge tubes or threaded tubes, and sometimes to mix the different primers or probes into the same tube, typically by a human operator using a pipette. The completion of the unequal number of separate aliquots of different primers in 96 wells, or the mixing of different primers in varying amounts, is a very tedious task itself. Because of the need to dispense or mix primers or probes in microliters, the skill requirements for the operator are high and it is often difficult to control the metering deviation or fluctuation to a small extent. Meanwhile, manual operation cannot completely avoid human errors, and is difficult to find when errors occur.

The laboratory pipetting workstations or nucleic acid extractors in the prior art can only realize the purpose of transferring certain liquid from one container to another container, and cannot realize the complex and precise requirements of split-charging and mixing of primers or probes. The measurement unit is usually in the milliliter level, the measurement precision is about 1%, the split charging or mixing of the primer or the probe is in the microliter level, the measurement precision requirement is higher, and the laboratory pipetting workstation or the nucleic acid extractor cannot meet the requirements of high precision and high stability of the micro-metering. Moreover, laboratory pipetting workstations or nucleic acid extractors are not capable of automatic collection of data modules nor of compatibility and coupling with DNA primer or probe synthesis systems.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, and aims to provide a device and a method for realizing two functions of split charging and mixing, comprising computer system control for realizing the functions, and a device for precisely split charging and mixing DNA primers or probes and a using method thereof.

The invention is realized by the following technical scheme: the accurate split charging and mixing device for the DNA primers or the probes mainly comprises a control cabinet, an executing mechanism, a working area part, a testing unit, a gun head frame, a working table plate and a fixed table plate; the actuating mechanism consists of an X-axis sliding table, a Y-axis sliding table, a Z-axis sliding table and a gun-axis sliding table; the X-axis sliding table is fixedly arranged on the positive direction surface of the fixed platen and is fixedly connected with the control cabinet; the working area component comprises 8 split charging target plates, 1 gun head plate, 1 stock solution plate, 1 liquid guiding target plate and 1 gun head separator, wherein the working table adopts a layout form with the stock solution plate as a center, the 8 split charging target plates are uniformly distributed on two sides of the stock solution plate in the X-axis direction, 4 split charging target plates in the positive direction are arranged in the negative direction, 4 split charging target plates in the negative direction are arranged in the positive direction of the Y-axis of the stock solution plate, and the liquid guiding target plates and the gun head separators are respectively arranged in the negative direction of the Y-axis of the stock solution plate.

The X-axis sliding table is connected with the Y-axis sliding table through a gantry type structure, an X-axis carriage is fixedly arranged on the upper surface of the X-axis sliding table, two upright posts are fixed on the X-axis carriage to support the Y-axis sliding table through bolts, the Z-axis sliding table is connected with the Y-axis sliding table through a side hoisting structure, a Y-axis carriage is fixedly arranged on the upper surface of the Y-axis sliding table, a Z-axis sliding table is fixedly arranged on the Y-axis carriage, and a gun-axis sliding table is arranged on the side surface of the Z-axis sliding table.

An internal controller, a direct-current power supply, a driver and a device driving motor are arranged in the control cabinet, and the device driving motor drives the X-axis slide plate, the Y-axis slide plate, the Z-axis slide plate and the gun-axis slide plate to move. The surface of the split charging target plate is provided with 12 multiplied by 5 hole sites, and the upper right corner of each hole site is provided with a semi-elliptical step bulge for fixing the tube cap of the centrifuge tube. The test unit is arranged on the upper surface of the workbench plate and is positioned in the positive direction of the Y axis of the stock solution plate.

The invention also discloses a use method of the accurate split charging and mixing device of the DNA primer or the probe, which comprises three steps of split charging or mixing requirement parameter obtaining step, medium extracting step from a designated pipe or a plate, pipetting step and medium quantitatively beating step into the designated pipe or the plate, wherein the specific operation process of each step comprises the following steps:

step one, obtaining split charging or mixing requirement parameters:

1) Selecting on-line operation or off-line operation with the DNA synthesizer at a computer control interface;

2) Selecting a split charging mode or a mixing mode;

3) Reading the demand data, namely the specific requirements of split charging or mixing;

when the accurate split charging and mixing device and the DNA synthesizer are in online operation, in the DNA synthesis digital management system, according to the requirement input of a customer order, the device adopts a digital module acquisition and management system compatible with the DNA synthesis digital management system, so as to realize direct introduction of required data;

when the accurate split charging and mixing device and the DNA synthesizer are operated offline, split charging or mixing demand information is directly established in a folder to be operated of a control computer of the device, wherein the information comprises initial position and volume of a split charging or mixing medium and position information to be transferred;

4) Converting the split charging or mixing requirement parameters into linkage operation instructions of all mechanisms of the device;

step two, extracting the medium from the specified tube or plate:

1) The main machine head unit is linked with the sliding table mechanisms in the X direction and the Y direction and moves to the upper part of the gun head frame;

2) The main machine head is fixed with a liquid-transferring gun head through the downward movement of the Z axis of the main machine head, and then is lifted to a proper height;

3) The main machine head unit with the gun head inserted is linked with the sliding table mechanisms in the X direction and the Y direction again, moves to the upper part of the stock solution plate, and enables the gun head to be inserted into the liquid in the corresponding hole site of the stock solution plate through the downward movement of the Z axis;

4) A motor on the main machine head drives a piston rod to extract and suck DNA liquid with a given dosage into the pipette head;

5) The main machine head Z moves upwards to separate the gun head from the stock solution and lifts the gun head to a proper height, so that the gun head cannot contact with a plate or a pipe on the operation table board in the moving process;

step three, pipetting and quantitatively beating medium into a designated tube or plate:

1) Moving the host head carrying the DNA liquid to the position above the corresponding centrifuge tube or centrifuge hole through the linkage of the sliding table mechanisms in the X direction and the Y direction;

2) The pipette gun heads are inserted into corresponding plate holes or centrifuge tubes through the downward movement of the Z shaft of the main machine head;

3) A motor on the host head drives a piston rod to push and throw out DNA liquid with given dosage into a corresponding plate hole or a centrifuge tube;

4) The main machine head Z moves upwards and is lifted to a proper height, so that preparation is made for subsequent movement;

5) The main machine head is linked with the sliding table mechanisms in the X direction and the Y direction and moves to the position of a gun head separator, the gun head separator clamps the gun head, the main machine head moves upwards along the Z axis, and the used gun head is dismounted;

6) And the main machine head moves to an initial position to finish the split charging or mixed pipetting operation of one primer or probe.

According to the requirement of split charging or mixing, the medium in the holes of several source plates needs to be split charging or mixing, and the second step and the third step are repeated for several times.

In medical and biological scientific research production, primers or probes with different base sequences and lengths are designed according to certain medical detection, diagnosis, research and development purposes, and finished products are stored in an orifice plate or a centrifuge tube after the procedures of molecular synthesis, ammonolysis, purification and the like. And then sub-packaging or mixing according to specific requirements and needs.

1. And (5) subpackaging:

the general DNA primer product is usually in a standard well plate, such as a 48-well plate or a 96-well plate, and each well may be a specific primer from different customers in different demands. According to the specific requirements of customers, each primer collected or temporarily stored in each well needs to be divided into a plurality of parts with determined dosages, packaged and stored in a centrifuge tube, which is called a plate-to-tube split charging process.

DNA probes or some finished primers subjected to special purification processes are usually collected and stored in larger centrifuge tubes, and are divided into a plurality of parts of defined doses according to the specific requirements of customers, and packaged and stored in smaller centrifuge tubes, which is called a tube-to-tube split charging process.

For subsequent mixing purposes, primers or probes buffered in the well plate or in larger centrifuge tubes need to be divided into several portions according to a certain dosage and portion, buffered in designated wells of a standard 96 well plate. The former is referred to as a "board-to-board" racking process, and the latter is referred to as a "tube-to-board" racking process.

2. Mixing:

for medical and biological research purposes, it is sometimes necessary to mix different primers or probes in 96-well plates or centrifuge tubes in a given dose. Similar to the dispensing process, the mixing process may also be a "plate-to-plate", "tube-to-plate", "plate-to-tube" or "tube-to-tube" mixing process. Typically, if more kinds of mixing need to be done at a time and sub-packaged again after mixing or the experiment is continued, the final mixing is more in 96-well plates; if there are only a few and no further sub-packaging is required after mixing, it can be mixed directly into a centrifuge tube.

DNA primer-primer, also known as primer. Is a small single-stranded DNA or RNA, and serves as a starting point for DNA replication, and the 3'-OH of the primer must be free because the nucleotide is synthesized as a diester chain on the 3' -OH of the primer, which serves as a starting point for extension of each polynucleotide strand during nucleic acid synthesis reaction.

DNA probes-Gene probes, i.e., nucleic acid probes (probes), are nucleic acid sequences (DNA or RNA) with a detectable label and known in sequence that are complementary to a gene of interest.

The 48-well plate or 96-well plate is widely used in the scientific research and production fields of biomedical science and the like, and is used as a cell culture vessel, a fluorescent quantitative PCR platform consumable or a nucleic acid collection temporary storage vessel and the like.

Centrifuge tube-tubular sample container, which may have sealing cover or gland for storing synthesized primer or probe.

The precise split charging and mixing device provided by the invention is similar to a pipetting workstation or a nucleic acid extractor for medical laboratories. The device and the method of the invention provide various innovative design schemes by combining the performance characteristics of the primer or probe materials, the special requirements of split charging or mixing, the control precision and the like, and realize functions and solutions which cannot be realized by a laboratory pipetting workstation or a nucleic acid extractor.

The beneficial effects of the invention are as follows: the invention provides a precise and reliable efficient engineering solution for a DNA primer or probe precise split charging and mixing device, which comprises the device and a using method thereof, so as to realize automatic precise split charging and mixing of the DNA primer or probe. By adopting the device and the using method, labor can be saved, the device can save the tedious manual liquid sucking and pumping process, an operator only needs to use the mobile U disk to guide the required operation into software, and the previous preparation work is saved. The safety and stability of the device for completing the work are far higher than those of the manual work, and errors can not occur due to the manual reasons. The precision of the device can reach within 2 percent, and the split charging operation of a split charging target plate can be completed within 30 minutes, so that the split charging and pipetting operations with high precision and wide range are realized. The device can realize full-automatic operation, and can be convenient for the automatic integration of the whole process in the later stage through the integration of the device.

The device adopts the plate type arrangement of eight target plates with brand new design, can realize most of the working requirements of DNA primer split charging, and does not need to suspend operations such as plate replacement and the like. The split charging of 96 primers can be completed at one time, so that the precision and the speed of split charging and pipetting of the primers are greatly improved, the preparation efficiency is improved, and the consistency and the reliability of the preparation are ensured. The invention can finish the accurate measurement from extraction to split charging or mixing of the primer or the probe. The movement process is stable and reliable, and the positioning is accurate. The accurate sub-packaging and mixing device has the advantages that: the functions of inserting gun heads, absorbing liquid, beating liquid, changing hole sites to beat liquid again and the like are realized, and five tubes can be split charged at most.

Drawings

FIG. 1 is a schematic diagram of the process of extracting media from a given tube/plate using the apparatus of the present invention;

FIG. 2 is a schematic perspective view of a dispensing and mixing device according to the present invention;

in fig. 2: 1-a control cabinet; 2-X axis sliding table; 3-Y axis sliding table; a 4-Z axis sliding table; 5-a gun shaft sliding table; 6-a test unit; 7-sub-packaging target plates; 8-gun head frame; 9-a stock solution plate; 10-a liquid guiding target plate; 11-gun head separator; 12-a work table; 13-fixing the bedplate; 14-hole site; 15-semi-elliptical step protrusions.

FIG. 3 is a schematic diagram of the hardware architecture of the control system of the racking and mixing apparatus of the present invention;

FIG. 4 is a schematic diagram of the connection and control principle of the actuator of the present invention;

FIG. 5 is a schematic perspective view of a dispensing target plate in the dispensing and mixing apparatus of the present invention;

wherein: the control cabinet holds all control parts, the test unit is used for realizing the test to the liquid filling amount of the device, the split charging target plate is used for holding the split charged primer, the gun head frame is used for holding the liquid-transferring gun head to be tested, and the stock solution plate is used for holding the original primer to be split charged.

Detailed Description

The invention is described in detail below with reference to the drawings and the detailed description.

As shown in fig. 1 to 5, an accurate packing and mixing device for DNA primers or probes is mainly composed of a control cabinet 1, an actuator, a working area part, a test unit 6, a gun head frame 8, a working platen 12 and a fixed platen 13; the actuating mechanism consists of an X-axis sliding table 2, a Y-axis sliding table 3, a Z-axis sliding table 4 and a gun-axis sliding table 5; the surface of the fixed platen 13 in the negative direction is fixedly connected with a control cabinet 1, and the X-axis sliding table 2 is fixedly arranged on the surface of the fixed platen 13 in the positive direction and is fixedly connected with the control cabinet 1; the working area component is fixedly connected to the working table plate 12 and comprises 8 split charging target plates 7, 1 gun head plate, 1 stock solution plate 9, 1 liquid guiding target plate 10 and 1 gun head separator 11, the working table plate 12 adopts a layout form taking the stock solution plate 9 as a center, the 8 split charging target plates 7 are uniformly distributed on two sides of the stock solution plate 9 in the X-axis direction, the 4 split charging target plates 7 in the positive direction and the 4 split charging target plates 7 in the negative direction are respectively arranged on the Y-axis positive direction of the stock solution plate 9, and the gun head frame 8 is respectively arranged on the Y-axis negative direction of the stock solution plate 9 by the liquid guiding target plate 10 and the gun head separator 11. The layout form taking the stock solution plate 9 as the center can improve the speed of split charging and liquid guiding and shorten the time of split charging and liquid guiding.

X axle slip table 2 adopts gantry structure to be connected with Y axle slip table 3, and the upper surface fixed of X axle slip table 2 is provided with the X axle carriage, holds up Y axle slip table 3 by two stand of bolt fastening on the X axle carriage, and Z axle slip table 4 adopts the structural style of side hoist and mount to be connected with Y axle slip table 3, is provided with the Y axle carriage at the upper surface fixed of Y axle slip table 3, fixedly on the Y axle carriage is provided with Z axle slip table 4, and rifle axle slip table 5 sets up the side at Z axle slip table 4. Thus, three degrees of freedom of linear motion are achieved.

An internal controller, a direct-current power supply, a driver and a device driving motor are arranged in the control cabinet 1, and the device driving motor drives the X-axis slide plate, the Y-axis slide plate, the Z-axis slide plate and the gun-axis slide plate to move. The surface of the split charging target plate is provided with 12×5 hole sites 14, and the upper right corner of each hole site 14 is provided with a semi-elliptical step bulge 15 for fixing the tube cap of the centrifuge tube. The test unit 6 is disposed on the upper surface of the table plate 12 in the positive Y-axis direction of the stock solution plate 9.

The main machine head piston rod unit of the device is driven by a stepping motor, and the stepping motor drives the screw rod to rotate to convert the rotary motion into the linear reciprocating motion of the piston rod, so that the liquid pumping and beating functions are realized. The lead of the screw rod is 2mm, and the linear reciprocating motion precision of the piston rod can reach 0.1mm. The whole motion unit consists of three ball screw linear sliding tables. The repeated positioning precision is below 0.02mm, and the stable and reliable work of the device in a high-precision state can be fully ensured.

The structure of the split charging and mixing device of the present invention is shown in fig. 2. First, X axle slip table 2 adopts gantry type structural style with Y axle slip table 3, and this structural style does not have fixed upper beam, can greatly save device space, makes things convenient for the work of going up liquid and getting liquid. The Z-axis sliding table 4 and the Y-axis sliding table 3 adopt a side hoisting structural form, so that the installation and maintenance of the Z-axis sliding table 4 are convenient. The gun shaft sliding table 5 is arranged on the side face of the Z-axis sliding table 4, and the gun shaft sliding table 5 adopts an open structure, so that the liquid takers with different volumes and gun heads thereof can be replaced conveniently. The device adopts two-layer bedplate forms, the lower floor is the fixed bedplate 13 of installation X axle slip table 2, and the upper strata is the work bedplate 12 of placing rifle headstock 8, former liquid board 9, partial shipment target board 7 and liquid guide target board 10 etc. work bedplate 12 is fixed, and the liquid of required operation can keep stationary state, guarantees the quality of putting of getting of liquid.

The control system hardware of the split charging and mixing device is shown in fig. 3, and is arranged in a device control cabinet 1 and mainly comprises a device internal controller, a direct current power supply, a driver and a device driving motor. The device internal controller is connected with the computer through an RJ45 network interface, receives split-charging liquid guiding data and instructions transmitted by the computer, analyzes the split-charging liquid guiding data and instructions, converts the split-charging liquid guiding data and instructions into a driver voltage driving signal, and drives the X-axis sliding plate, the Y-axis sliding plate, the Z-axis sliding plate and the gun-axis sliding plate to move through a device driving motor after the driver voltage driving signal is amplified. The target interpolation and movement of the gun head frame 8, the stock solution plate 9, the split charging target plate 7 and the liquid guiding target plate 10 are realized through the positioning data of the gun head frame 8, the stock solution plate 9, the split charging target plate 7 and the liquid guiding target plate 10, and the gun head is driven to move to the appointed positions of the gun head frame 8, the stock solution plate 9, the split charging target plate 7 and the liquid guiding target plate 10 for liquid absorption and liquid discharge, so that the device functions of split charging and liquid guiding are realized.

The actuating mechanism consists of an X-axis sliding table 2, a Y-axis sliding table 3, a Z-axis sliding table 4 and a gun-axis sliding table 5. The control mode of each sliding table is the same and is the same as an independent control unit. The control principle and the operation logic of the execution mechanism are as follows:

1) An operator firstly makes split charging liquid guide data files, and saves the data as excel data files;

2) Operating device operation software, importing split charging liquid guide data into the operation software, and definitely designating the device as a split charging working mode or a liquid guide working mode;

3) The device operation software automatically reads the split charging liquid guide data file through the RJ45 network connecting line connected with the computer and the controller in the device;

4) The device operation software converts the split charging liquid guide data into control instructions and sends the control instructions to the internal controller of the device through a network protocol;

5) The internal controller of the device converts the control instruction into interpolation motion voltage driving signals of an X axis, a Y axis and a Z axis and sends the signals to a driver;

6) The driver amplifies and distributes the voltage signals and then sends the voltage signals to drive motors of an X axis, a Y axis and a Z axis;

7) The motors of the X axis, the Y axis and the Z axis drive the gun head to move to the appointed positions of the gun head frame 8, the stock solution plate 9, the split charging target plate 7 and the liquid guiding target plate 10;

8) The gun shaft motor drives the gun shaft slide plate to move according to the liquid amount command, and liquid with specified liquid amount is sucked and placed;

9) And after the motors of the X axis, the Y axis, the Z axis and the gun axis receive the stop instruction, stopping the operation of the motors, and completing the split charging and liquid guiding work.

Fig. 4 is a control schematic diagram of the sliding table: the motor is linked with the screw rod through a coupler, and the carriage is fixed on a nut on the screw rod. The motor can transmit the rotation motion to the carriage, so that the carriage can finally complete the linear reciprocating motion, and the principle of the sliding table in each direction is the same.

As shown in fig. 2, the device is independently provided with a calibration position, the right hole is the gun head position of the calibration position, the middle hole is the raw liquid hole, and the left hole is the calibration target hole. In the calibration interface drop down menu, calibration positions can be corrected, where the operator only needs to calibrate the gun head position.

As shown in fig. 5, the split charging target plate 7 adopts a novel design of a centrifuge tube frame plate, the centrifuge tube frame is made of aluminum materials, the surface of the centrifuge tube frame comprises 12×5 hole sites 14, and a convex semi-elliptical step is arranged at the upper right corner of each hole site 14 for fixing a centrifuge tube cover.

The application method of the device can be divided into the steps of obtaining the parameters of the split charging/mixing requirements, extracting the medium (primer or probe) from the designated tube or plate, pipetting, quantitatively beating the medium into the designated tube/plate and the like, and the specific operation process of each step is as follows:

step one: obtaining split charging or mixing requirement parameters:

the aim of the step is to be compatible with a general data module, acquire the required parameters to be split-charged or mixed in a convenient way and convert the required parameters into operation control instructions of the device.

1.1, selecting online operation or offline operation with a DNA synthesizer on a computer control interface;

1.2, selecting a split charging mode or a mixing mode;

1.3, reading the required data, namely the specific requirements of split charging or mixing;

1.3.1, when the device is in online operation with a DNA synthesizer, the DNA synthesis digitalized management system is input according to the requirement of a customer order, and the device adopts a digital module acquisition and management system compatible with the device, so that the direct importing of the requirement data can be realized;

1.3.2, when the device and the DNA synthesizer are operated offline, directly establishing split charging or mixing requirement information in a folder to be operated of a control computer of the device, wherein the split charging or mixing requirement information comprises information such as initial positions (holes and pipes) of a to-be-split charging or mixing medium, volumes, positions (holes and pipes) to be transferred and the like;

and 1.4, converting the split charging or mixing requirement parameters into linkage operation instructions of all mechanisms of the device.

Step two: extraction of media from designated tubes/plates:

the method aims at accurately extracting a certain volume of primers or probes from a designated plate hole or a centrifuge tube by three-dimensional linkage of each motion mechanism according to the required parameters to be split-packed or mixed.

2.1, step 1 in fig. 1: the main machine head unit is linked with the sliding table mechanisms in the X direction and the Y direction and moves to the position above the gun head frame 8; step 2 in fig. 1: the main machine head is fixed with a liquid-transferring gun head through the downward movement of the Z axis of the main machine head, and then is lifted to a proper height; step 3 in fig. 1: the main machine head unit with the gun head inserted is linked with the sliding table mechanisms in the X direction and the Y direction again and moves to the upper part of the stock solution plate 9 or the pipe, and the gun head is inserted into the liquid in the corresponding hole site 14 of the stock solution plate 9/the pipe through the downward movement of the Z axis; step 4 in fig. 1: the used gun head is detached.

2.2, a motor on the main machine head drives a piston rod to extract and suck DNA liquid with a given dosage into the pipette head.

2.3, the main machine head Z moves upwards to separate the gun head from the stock solution and lifts to a proper height, so that the gun head can not contact with the plate/tube on the operating table in the moving process.

Step three: pipetting and quantifying out medium into designated tubes/plates:

the main purpose of this step is to move the extracted DNA primer or probe liquid to the designated plate well or centrifuge tube location and to achieve accurate ejection of the medium required dose.

And 3.1, moving the host head carrying the DNA liquid to the upper part of a corresponding centrifuge tube or a centrifuge hole through the linkage of the sliding table mechanisms in the X direction and the Y direction.

3.2, inserting the pipette tips into corresponding plate holes or centrifuge tubes through the downward movement of the Z axis of the main machine head.

And 3.3, driving a piston rod by a motor on the host head, and pushing out the DNA liquid with a given dosage into a corresponding plate hole or a centrifuge tube.

3.4, the main machine head Z moves upwards and is lifted to a proper height, so that the following movement is prepared.

If the DNA primer or probe liquid is required to be distributed into a plurality of tubes/holes, the DNA liquid pipetting and distribution and extraction of each designated tube/hole are completed according to the same principle of the steps 3.1 to 3.4, thereby achieving the purposes of one-time extraction and porous distribution.

3.5, as shown in fig. 1, the main machine head is linked with the sliding table mechanisms in the X direction and the Y direction and moves to the position of the gun head separator 11, the gun head separator 11 clamps the gun head, and the main machine head moves upwards along the Z axis. As shown in step 4 of FIG. 1, the used tips are removed, and each primer or probe is extracted, pipetted and ejected with only one tip at a time to avoid cross contamination.

And 3.6, the host head moves to an initial position to finish the split charging or mixed pipetting operation of one primer or probe.

According to the requirement of split charging or mixing, the primers/probes in the source plate holes/tubes need to be split charging or mixing, and the second step and the third step are repeated for several times. The device can automatically connect and complete all operations according to the complete parameters obtained in the first step, no pause occurs in the middle, and the operation can be continued by the button pause and the button recovery during the reuse unless manual intervention is required for special reasons.

Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. An accurate partial shipment and mixing arrangement of DNA primer or probe, its characterized in that: the precise split charging and mixing device mainly comprises a control cabinet, an actuating mechanism, a working area component, a testing unit, a gun head frame, a working table plate and a fixed table plate; the actuating mechanism consists of an X-axis sliding table, a Y-axis sliding table, a Z-axis sliding table and a gun-axis sliding table; the control cabinet is fixedly connected to the upper surface of the negative direction of the fixed platen, and the X-axis sliding table is fixedly arranged on the upper surface of the positive direction of the fixed platen and is fixedly connected with the control cabinet; the working area component comprises 8 split charging target plates, 1 gun head frame, 1 stock solution plate, 1 liquid guiding target plate and 1 gun head separator, wherein the working area component adopts a layout form taking the stock solution plate as a center, the 8 split charging target plates are uniformly distributed on two sides of the X-axis direction of the stock solution plate, the 4 split charging target plates are in positive direction, the 4 split charging target plates are in negative direction, the gun head frame is arranged in the Y-axis positive direction of the stock solution plate, the liquid guiding target plates and the gun head separator are respectively arranged in the Y-axis negative direction of the stock solution plate, and the gun head plate, the stock solution plate, the liquid guiding target plate and the gun head separator are positioned in the area between the 4 split charging target plates in positive direction and the 4 split charging target plates in negative direction, and the gun head frame, the stock solution plate, the liquid guiding target plate and the gun head separator are sequentially arranged along the Y-axis positive direction to the Y-axis negative direction; the accurate partial shipment and mixing arrangement has partial shipment mode and the mixed mode to DNA primer or probe just accurate partial shipment and mixing arrangement can with DNA synthesizer online operation or off-line operation.

2. The precise packing and mixing device for DNA primers or probes according to claim 1, wherein: the X-axis sliding table is connected with the Y-axis sliding table through a gantry structure, an X-axis carriage is fixedly arranged on the upper surface of the X-axis sliding table, two upright posts are fixed on the X-axis carriage to support the Y-axis sliding table, the Z-axis sliding table is connected with the Y-axis sliding table through a side hoisting structure, a Y-axis carriage is fixedly arranged on the upper surface of the Y-axis sliding table, a Z-axis sliding table is fixedly arranged on the Y-axis carriage, and the gun-axis sliding table is arranged on the side surface of the Z-axis sliding table.

3. The precise packing and mixing device for DNA primers or probes according to claim 1 or 2, wherein: an internal controller, a direct-current power supply, a driver and a device driving motor are arranged in the control cabinet, and the device driving motor drives the X-axis slide plate, the Y-axis slide plate, the Z-axis slide plate and the gun-axis slide plate to move.

4. The precise packing and mixing device for DNA primers or probes according to claim 1 or 2, wherein: the surface of the split charging target plate is provided with 12 multiplied by 5 hole sites, and the upper right corner of each hole site is provided with a semi-elliptical step bulge for fixing the tube cap of the centrifuge tube.

5. The precise packing and mixing device for DNA primers or probes according to claim 1 or 2, wherein: the testing unit is arranged on the upper surface of the workbench plate and located in the positive direction of the Y axis of the stock solution plate.

6. A method of using the DNA primer or probe precision dispensing and mixing device of any one of claims 1 to 5, characterized in that: the method comprises three steps of sub-packaging or mixing requirement parameter obtaining, medium extracting from a designated pipe or plate and pipetting and quantitatively beating medium into the designated pipe or plate, wherein the specific operation process of each step comprises the following steps:

step one, obtaining split charging or mixing requirement parameters:

2) Selecting a split charging mode or a mixing mode;

step two, extracting the medium from the specified tube or plate:

7. The method for using a precise packing and mixing device for DNA primers or probes according to claim 6, wherein the method comprises the following steps: in the first step, when the accurate split charging and mixing device and the DNA synthesizer are in online operation, in the DNA synthesis digital management system, the device adopts a digital module acquisition and management system compatible with the device according to the demand input of a customer order, so as to realize direct introduction of demand data.

8. The method for using a precise packing and mixing device for DNA primers or probes according to claim 6, wherein the method comprises the following steps: in the first step, when the accurate sub-packaging and mixing device and the DNA synthesizer are in offline operation, sub-packaging or mixing requirement information is directly established in a folder to be operated by a control computer of the device, wherein the information comprises initial position and volume of a sub-packaging or mixing medium and position information to be transferred.

9. The method for using a precise packing and mixing device for DNA primers or probes according to claim 6, wherein the method comprises the following steps: according to the requirement of sub-packaging or mixing, the media in the holes of the source plates are required to be sub-packaged or mixed, and the second step and the third step are repeated for several times.