Automatic cell transformation workstation and working method thereof

Abstract

The invention discloses an automatic cell transformation work station and a working method thereof, and belongs to the technical field of molecular biology experiment automation, the automatic cell transformation work station comprises a work station box body, the work station box body is internally provided with a constant temperature oscillation mechanism, a metal bath mechanism, two water bath mechanisms, a pipetting mechanism and a transfer mechanism; the pipetting mechanism is used for adding each sample into the deep hole plate; the constant-temperature oscillation mechanism is used for performing oscillation recovery culture on a sample in the deep-hole plate at a constant temperature; the metal bath mechanism is used for carrying out constant-temperature heat preservation on a sample in the PCR plate; the two water bath mechanisms are respectively used for carrying out cold bath treatment and heat shock treatment on a sample in the deep hole plate; and the transfer mechanism is used for transferring the deep-hole plate. The device integrates constant-temperature oscillation, metal bath heat preservation, water bath cold shock and water bath heat shock, the automation degree of an experiment is improved, human intervention is reduced, the pollution risk is reduced, and the experiment efficiency is improved.

Description

Technical Field

[0001] The invention relates to the technical field of molecular biological experiment automation, in particular to an automated cell transformation workstation and a working method thereof. Background Art

[0002] Transformation refers to the phenomenon that a cell of a certain genotype absorbs exogenous DNA / RNA from the surrounding medium, causing its genotype and phenotype to change accordingly. Bacterial cell transformation is a very common molecular experimental method, including chemical transformation and electroporation transformation methods, and is widely used in life science research.

[0003] In the most commonly used chemical transformation method, a series of operations are required to mix the target competent bacterial cells with the target DNA / RNA samples, heat shock, cold bath, constant temperature culture, etc. The factors that affect the transformation efficiency of the competent bacterial cells include the quality of the reagents themselves, the time for the competent cells to melt, the sample mixing method and static time, the temperature and time of the heat shock cold bath, and the stability and speed of the sample's multiple transfers in various devices. When there are many samples, it will take a long time for a single person to transform bacterial cells, and it is difficult to ensure that the transformation efficiency of each sample is basically the same. In the process, the risk of contamination between samples will also increase. Multiple people operating cannot avoid the risk of contamination between samples, and it also increases labor costs.

[0004] To this end, an automated cell transformation workstation and a working method thereof are proposed. Summary of the invention

[0005] The object of the present invention is to provide an automated cell transformation workstation and a working method thereof, aiming to solve or improve at least one of the above-mentioned technical problems.

[0006] To achieve the above object, the present invention provides the following solutions: The present invention provides an automated cell transformation workstation, comprising a workstation box, wherein a constant temperature oscillation mechanism, a metal bath mechanism, two water bath mechanisms, a liquid transfer mechanism and a transport mechanism are arranged in the workstation box;

[0007] The pipetting mechanism is used to add each sample into the deep well plate;

[0008] The constant temperature shaking mechanism is used to shake and resuscitate the samples in the deep well plate at a constant temperature;

[0009] The metal bath mechanism is used to keep the samples in the PCR plate at a constant temperature;

[0010] The two water bath mechanisms are used to perform cold bath treatment and heat shock treatment on the samples in the deep well plate respectively;

[0011] The transport mechanism is used for transporting the deep well plate.

[0012] Optionally, the water bath mechanism includes a water bath disposed in the workstation box, the water bath is detachably connected to the deep well plate, a liquid supply assembly is disposed in the workstation box, the liquid supply assembly is connected to the water bath, a cooling and heating assembly is disposed on the water bath, and a temperature sensor is fixedly connected in the water bath.

[0013] Optionally, the cooling and heating component includes a semiconductor cooling and heating plate fixedly connected to the bottom of the water bath, a copper heat pipe is fixedly connected to the bottom of the water bath, and the semiconductor cooling and heating plate is in contact with the copper heat pipe and the outer bottom wall of the water bath.

[0014] Optionally, a water-cooled copper bus is further provided at one end of the semiconductor refrigeration and heating plate away from the water bath, and the water-cooled copper bus includes a copper shell fixedly connected to the semiconductor refrigeration and heating plate, and a first circulation pipe and a second circulation pipe are fixedly connected and connected to the copper shell, the first circulation pipe is fixedly connected and connected to a water cooling box, a water pump is fixedly connected in the water cooling box, and the water outlet end of the water pump is fixedly connected and connected to the second circulation pipe.

[0015] Optionally, a plurality of spring sheets are circumferentially fixed to the outer wall of the water bath, and the deep hole plate is clamped between the spring sheets.

[0016] Optionally, the liquid supply assembly includes a liquid bottle fixedly connected to the workstation box, a liquid inlet is provided on the water bath, the liquid bottle is fixedly connected and connected to the water inlet of the peristaltic pump through a first liquid tube, and the water outlet of the peristaltic pump is fixedly connected and connected to the liquid inlet through a second liquid tube.

[0017] Optionally, the water bath outer cover is provided with a heat insulation cover with an opening at the top, a heat insulation cover is provided on the top of the heat insulation cover, the heat insulation cover is fixedly connected to the workstation box, and the heat insulation cover is covered on the top of the deep well plate.

[0018] Optionally, a first support plate is fixedly connected in the workstation box, and the water bath, the constant temperature oscillation mechanism, and the metal bath mechanism are all fixedly connected to the first support plate; the pipetting mechanism includes a moving component fixedly connected to the workstation box, and an electric multi-channel pipette is fixedly connected to the output end of the moving component, and the moving component drives the electric multi-channel pipette to move along the X-axis, Y-axis and Z-axis directions.

[0019] Optionally, an air filter and an ultraviolet lamp are fixedly connected to the inner top wall of the workstation box.

[0020] A method for automated cell transformation is also provided, comprising the following steps:

[0021] Step 1: Pre-start

[0022] Start the constant temperature oscillation mechanism, the metal bath mechanism and the two water bath mechanisms, set the preheating and precooling temperatures and make them reach the set values; at the same time, disinfect the workstation;

[0023] Step 2: Place the actual consumables in the designated location

[0024] Place the 1000uL pipette tip, 20uL pipette tip, PCR plate containing DNA samples, deep-well plate containing competent cells, SOC medium, heat-insulating cover, and sponge pad in designated locations respectively;

[0025] Step 3: Lysis of competent cells

[0026] The transfer mechanism transfers the insulation cover to the deep-well plate and leaves it to stand for 15 minutes to allow the competent cells to slowly dissolve in the pre-cooled water bath mechanism;

[0027] Step 4: Sample mixing

[0028] After the standing is completed, the transport mechanism moves the insulation cover away from the deep-well plate and returns it to its original position. The pipetting mechanism absorbs all DNA samples from the PCR plate and adds them to the deep-well plate. The samples are mixed with the competent cells in the deep-well plate by in-situ aspiration and release. The insulation cover is then placed on the deep-well plate by the transport mechanism and allowed to stand for 30 minutes to allow the DNA samples to be in contact with the competent cells and stabilize.

[0029] Step 5: Heat shock treatment

[0030] After the static state is completed, the transport mechanism quickly transports the deep-well plate to another preheated water bath mechanism for heat shock treatment;

[0031] Step 6: Cold Bath Treatment

[0032] After the heat shock is completed, the transport mechanism quickly transfers the deep-well plate back to the pre-cooled water bath mechanism for cold bath standing;

[0033] Step 7: Adding culture medium

[0034] The transport mechanism removes the heat-insulating cover on the deep-well plate, and the pipetting mechanism draws the required amount of SOC medium from the reservoir and adds it to the deep-well plate;

[0035] Step 8: Constant temperature shaking culture

[0036] The transport mechanism transfers the deep-well plate to the constant temperature shaking mechanism, and then transports the insulation cover back to cover the deep-well plate again, and shakes the deep-well plate at the constant temperature for 1 hour to revive the transformed competent cells.

[0037] The present invention discloses the following technical effects: the present application integrates constant temperature oscillation, metal bath insulation, water bath cold shock and water bath heat shock, wherein each sample is automatically added to a deep well plate through a pipetting mechanism, thereby avoiding human errors and improving the accuracy of experimental results; the deep well plate is automatically transported in each mechanism through a transport mechanism, thereby improving the degree of automation of the experiment, reducing human intervention, reducing the risk of contamination and improving experimental efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] The drawings constituting a part of the present application are used to provide a further understanding of the present application. The illustrative embodiments and descriptions of the present application are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0039] Figure 1 It is a schematic diagram of the internal structure of the workstation box of the present invention;

[0040] Figure 2 It is an exploded view of the water bath mechanism in the present invention;

[0041] Figure 3 It is a schematic diagram of the structure of the semiconductor refrigeration and heating plate and the copper heat conducting pipe in the present invention;

[0042] Figure 4 It is a schematic diagram of the structure of the temperature sensor in the present invention;

[0043] Figure 5 It is a positional relationship diagram of various consumables and various structures in the present invention.

[0044] In the figure: 1. workstation box; 2. constant temperature oscillation mechanism; 3. metal bath mechanism; 4. water bath mechanism; 5. water bath; 6. deep well plate; 7. semiconductor refrigeration and heating plate; 8. copper heat pipe; 9. copper shell; 10. first circulation pipe; 11. second circulation pipe; 12. water cooling box; 13. liquid bottle; 14. peristaltic pump; 15. heat insulation cover; 16. heat insulation cover; 17. first support plate; 18. first linear motion module; 19. second linear motion module; 20. third linear motion module; 21. electric multi-channel pipette; 22. air filter; 23. ultraviolet lamp; 25. temperature sensor. DETAILED DESCRIPTION

[0045] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0046] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0047] Reference Figure 1-Figure 4 The present invention provides an automated cell transformation workstation, comprising a workstation housing 1, wherein a constant temperature oscillation mechanism 2, a metal bath mechanism 3, two water bath mechanisms 4, a liquid transfer mechanism and a transport mechanism are arranged in the workstation housing 1;

[0048] The pipetting mechanism is used to add each sample into the deep well plate 6;

[0049] The constant temperature shaking mechanism 2 is used to shake and resuscitate the sample in the deep well plate 6 at a constant temperature;

[0050] The metal bath mechanism 3 is used to keep the samples in the PCR plate at a constant temperature;

[0051] The two water bath mechanisms 4 are used to perform cold bath treatment and heat shock treatment on the samples in the deep well plate 6 respectively;

[0052] The transport mechanism is used to transport the deep well plate 6 .

[0053] In this embodiment, the transport mechanism includes a robotic arm, a precision positioning sensor and a motion controller. The mechanical gripper on the robotic arm holds the deep-well plate 6, and the precise positioning sensor is used to accurately position the deep-well plate 6, and the controller controls the movement of the robotic arm, thereby performing a transport operation on the deep-well plate 6.

[0054] The constant temperature oscillation mechanism 2 includes a heating element, a temperature sensor, an oscillation motor and a controller, etc. The deep well plate 6 is placed in the bracket of the constant temperature oscillation mechanism 2, so as to facilitate oscillation at a constant temperature;

[0055] The present application integrates constant temperature oscillation, metal bath insulation, water bath cold shock, and water bath hot shock. Each sample is automatically added to the deep well plate 6 through a pipetting mechanism, thereby avoiding human errors and improving the accuracy of the experimental results. The deep well plate 6 is automatically transported in various mechanisms through a transport mechanism, thereby improving the degree of automation of the experiment, reducing human intervention, reducing the risk of contamination and improving experimental efficiency.

[0056] A further optimized solution is that the water bath mechanism 4 includes a water bath 5 arranged in the workstation box 1, the water bath 5 is detachably connected to the deep well plate 6, a liquid supply component is arranged in the workstation box 1, the liquid supply component is connected to the water bath 5, a cooling and heating component is arranged on the water bath 5, and a temperature sensor 25 is fixedly connected in the water bath 5.

[0057] According to a further optimized solution, a plurality of spring pieces are fixedly connected to the outer wall of the water bath 5 in the circumferential direction, and the deep hole plate 6 is sandwiched between the plurality of spring pieces.

[0058] The water bath 5 can be adapted to the deep-hole plate 6, and spring clips are arranged around it. After the deep-hole plate 6 is placed in the water bath 5, these spring clips will undergo elastic deformation and fit tightly to the side wall of the semi-skirt of the deep-hole plate 6, thereby ensuring the stability and positioning accuracy of the deep-hole plate 6 during the water bath process and improving the water bath effect.

[0059] A further optimized solution is that the cooling and heating component includes a semiconductor cooling and heating plate 7 fixedly connected to the bottom of the water bath 5, a copper heat pipe 8 is fixedly connected to the bottom of the water bath 5, and the semiconductor cooling and heating plate 7 is in contact with the copper heat pipe 8 and the outer bottom wall of the water bath 5.

[0060] A further optimized solution is that the liquid supply component includes a liquid bottle 13 fixedly connected to the workstation box 1, a liquid inlet is opened on the water bath 5, the liquid bottle 13 is fixedly connected and connected to the water inlet of the peristaltic pump 14 through a first liquid tube, and the water outlet of the peristaltic pump 14 is fixedly connected and connected to the liquid inlet through a second liquid tube.

[0061] During operation, the peristaltic pump 14 starts to work, and it pumps the coolant in the liquid bottle 13 into the water bath 5 through the first liquid pipe, so that the bottom of the deep well plate 6 is completely immersed in the coolant, thereby achieving effective heat exchange; the semiconductor refrigeration and heating plate 7 works to generate cooling or heating, wherein the coolant in the water bath 5 (T5 position) is cooled, and the coolant in the water bath 5 (T6 position) is heated, thereby cooling or heating the sample in the deep well plate 6 in the process; by setting the copper heat pipe 8, the heat exchange effect between the water bath 5 and the semiconductor refrigeration and heating plate 7 is improved.

[0062] A further optimization scheme is that a water-cooled copper busbar is provided at one end of the semiconductor refrigeration and heating plate 7 away from the water bath 5, and the water-cooled copper busbar includes a copper shell 9 fixedly connected to the semiconductor refrigeration and heating plate 7, and a first circulation pipe 10 and a second circulation pipe 11 are fixedly connected and connected to the copper shell 9, and the first circulation pipe 10 is fixedly connected and connected to a water cooling box 12, and a water pump is fixedly connected in the water cooling box 12, and a water outlet end of the water pump is fixedly connected and connected to the second circulation pipe 11.

[0063] There is heat dissipation coolant in the water cooling box 12. In order to maintain the continuous cooling effect of the semiconductor refrigeration and heating plate 7, the copper shell 9 below is close to the back of the semiconductor refrigeration and heating plate 7, and the heat generated by the semiconductor refrigeration and heating plate 7 is taken away by the continuous circulation of the heat dissipation coolant; this heat dissipation process ensures the continuous and efficient operation of the semiconductor refrigeration and heating plate 7, thereby ensuring the temperature control accuracy of the sample.

[0064] The water bath mechanism 4 in the present application achieves precise temperature control of the sample consumables through precise positioning of the deep well plate 6, efficient coolant circulation, and stable refrigeration and heating and heat dissipation mechanism during the refrigeration process.

[0065] According to a further optimization scheme, the outer cover of the water bath 5 is provided with a heat insulating cover 15 with an opening at the top, a heat insulating cover 16 is provided on the top of the heat insulating cover 15 , the heat insulating cover 15 is fixedly connected to the workstation box 1 , and the heat insulating cover 16 covers the top of the deep well plate 6 .

[0066] A further optimized solution is that a first support plate 17 is fixedly connected to the workstation box 1, and the water bath 5, the constant temperature oscillation mechanism 2, and the metal bath mechanism 3 are all fixedly connected to the first support plate 17; the pipetting mechanism includes a motion component fixedly connected to the workstation box 1, and an electric multi-channel pipette gun 21 is fixedly connected to the output end of the motion component, and the motion component drives the electric multi-channel pipette gun 21 to move along the X-axis, Y-axis and Z-axis directions.

[0067] The motion component includes a first linear motion module 18 fixedly connected in the workstation box 1, the moving end of the first linear motion module 18 is fixedly connected to the second linear motion module 19, the moving end of the second linear motion module 19 is fixedly connected to the third linear motion module 20, the moving end of the third linear motion module 20 is fixedly connected to the electric multi-channel pipette 21, the first linear motion module 18, the second linear motion module 19, and the third linear motion module 20 move along the X-axis, Y-axis, and Z-axis directions respectively, and the first linear motion module 18, the second linear motion module 19, and the third linear motion module 20 include but are not limited to linear motors, hydraulic cylinders, cylinders and the like.

[0068] According to a further optimized solution, an air filter 22 and an ultraviolet lamp 23 are fixedly connected to the inner top wall of the workstation box 1 .

[0069] In order to further enhance the cleanliness and safety inside the workstation box 1, an ultraviolet lamp 23 is also provided; after daily work is completed, the control system controls the ultraviolet lamp 23 to automatically turn on for disinfection without manual intervention.

[0070] The air filter 22 filters the external air to remove dust, particles, bacteria, viruses and other pollutants therein, and blows the filtered clean air into the workstation box 1. The workstation box 1 is provided with an air outlet for air circulation; the clean air forms a certain airflow circulation inside the workstation box 1, blowing harmful substances such as microorganisms and aerosols inside the equipment out of the working area, thereby maintaining the cleanliness of the equipment working area. This airflow circulation helps to reduce the accumulation of pollutants inside the equipment and the risk of cross contamination.

[0071] A method for automated cell transformation is also provided, comprising the following steps:

[0072] Step 1: Pre-start

[0073] Start the constant temperature oscillation mechanism 2 and the two water bath mechanisms 4, set the preheating and precooling temperatures and make them reach the set values; and disinfect the workstation at the same time;

[0074] Step 2: Place the actual consumables in the designated location

[0075] Place the 1000uL pipette tip, 20uL pipette tip, PCR plate containing DNA samples, deep-well plate 6 containing competent cells, SOC culture medium, heat-insulating cover 16, and sponge pad in designated positions respectively;

[0076] Step 3: Lysis of competent cells

[0077] The transport mechanism transfers the heat-insulating cover 16 to cover the deep-well plate 6, and leaves it to stand for 15 minutes to allow the competent cells to slowly dissolve in the pre-cooled water bath mechanism 4;

[0078] Step 4: Sample mixing

[0079] After the standing is completed, the transport mechanism moves the heat-insulating cover 16 away from the deep-well plate 6 and returns it to its original position. The pipetting mechanism absorbs all DNA samples from the PCR plate and adds them to the deep-well plate 6. The samples are mixed with the competent cells in the deep-well plate 6 by in-situ liquid aspiration and liquid discharge. The heat-insulating cover 16 is then covered on the deep-well plate 6 by the transport mechanism and allowed to stand for 30 minutes to allow the DNA samples to be in contact with the competent cells and to be stable.

[0080] Step 5: Heat shock treatment

[0081] After the static state is completed, the transport mechanism quickly transports the deep well plate 6 to another preheated water bath mechanism 4 for heat shock treatment;

[0082] Step 6: Cold Bath Treatment

[0083] After the heat shock is completed, the transport mechanism quickly transfers the deep well plate 6 back to the pre-cooled water bath mechanism 4 for cooling and standing;

[0084] Step 7: Adding culture medium

[0085] The transport mechanism removes the heat-insulating cover on the deep-well plate 6, and the pipetting mechanism draws a required amount of SOC culture medium from the reservoir and adds it into the deep-well plate 6;

[0086] Step 8: Constant temperature shaking culture

[0087] The transport mechanism transfers the deep-well plate 6 to the constant temperature shaking mechanism 2, and then transports the heat-insulating cover back to cover the deep-well plate 6 again, and shakes the deep-well plate 6 at the constant temperature for 1 hour to revive the transformed competent cells.

[0088] The specific operations are as follows:

[0089] Step 1: Pre-start

[0090] Start the constant temperature oscillation mechanism 2, the metal bath mechanism 3, and the two water bath mechanisms 4, set the preheating and precooling temperatures as shown in Table 1 and make them reach the set values, and turn on the purple light 23 to disinfect the workstation:

[0091] Table 1

[0092]

[0093] Step 2: Place the actual consumables as shown in Table 2. Figure 5 The specified location shown;

[0094] Table 2

[0095] Location Consumables and reagents T1 1000uL pipette tip T2 20uL pipette tip T4 PCR plate containing 10uL of enzyme-linked product sample T5 Deep well plate containing 100uL competent cells T7 SOC medium T9 Insulation cover T10 Sponge pad

[0096] Step 3: Lysis of competent cells

[0097] The transport mechanism moves the insulation cover to the T5 position and covers the deep-well plate 6, and leaves it to stand for 15 minutes to allow the competent cells to slowly dissolve in the pre-cooled water bath mechanism 4;

[0098] Step 4: Sample mixing

[0099] The transport mechanism moves the heat-insulating cover on the deep-well plate 6 back to the T9 position, and the pipetting mechanism absorbs all enzyme-linked product samples (10uL) from the PCR plate at the T4 position, and adds them to the deep-well plate 6 at the T5 position, and mixes them with the competent cells in the deep-well plate 6 by slowly aspirating and discharging the liquid twice in situ, and then the heat-insulating cover is moved from the T9 position back to the T5 position by the transport mechanism to cover the deep-well plate, and then allowed to stand for 30 minutes to allow the enzyme-linked product to be in contact with the competent cells and stabilize;

[0100] Step 5: Heat shock treatment

[0101] After standing for 30 minutes, the transport mechanism quickly transports the deep well plate 6 at position T5 to the preheated water bath mechanism 4 at position T6 for a 90 s heat shock treatment;

[0102] Step 6: Cold Bath Treatment

[0103] After the heat shock is completed, the transport mechanism quickly transfers the deep well plate 6 at position T6 back to the pre-cooled water bath mechanism 4 at position T5 for a cold bath and allows it to stand for 5 minutes;

[0104] Step 7: Adding culture medium

[0105] The transport mechanism moves the heat-insulating cover on the deep-well plate 6 at position T5 to position T9 and places it there. The pipetting mechanism draws 400uL of SOC culture medium from the reservoir at position T7 and adds it to the deep-well plate 6 at position T5.

[0106] Step 8: Constant temperature shaking culture

[0107] The transport mechanism transfers the deep-well plate 6 at position T5 to the constant temperature shaking mechanism 2, and then transports the insulation cover back from position T9 to cover the deep-well plate 6 again, and shakes at a constant temperature of 500 rpm for 1 hour to revive the transformed competent cells. In the above, the pipetting mechanism will automatically discard the used waste pipette tip into the T8 waste bucket.

[0108] The equipment of the present invention was used to perform high-throughput automated plasmid transformation as an experimental group, and the transformation was performed manually as a control group. The experimental conditions of the two groups were as similar as possible. The heat shock temperature was set to 42°C and the duration was 90 seconds by manual operation of the external equipment. The ice bath was placed on ice (0°C) for 30 minutes and 5 minutes respectively. The constant temperature shaking culture conditions were 37°C and 180rpm. 250uL of the finally obtained transformed competent cells was evenly coated on a culture dish containing resistance. The test results are shown in Table 3:

[0109] Table 3

[0110] index Experimental group (automation - 1 round) Control group (manual - 1 person) Flux 96 96 Time (end of recovery) About 2 hours About 3 hours Conversion success rate 100％ 100％ Number of spots normal Individual samples are too few

[0111] The experimental results show that the use of the present invention can shorten the experimental time. Although the final conversion success rate of the experimental group and the control group is 100%, the number of spots in some samples of the control group is relatively small, which may be caused by improper manual operation or unstable manual operation time control, resulting in poor conversion effect of some samples and high error rate. The use of the present invention basically does not require manual intervention, further saving labor costs.

[0112] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside" and "outside" etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0113] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. An automated cell transformation workstation, characterized in that: It comprises a workstation box (1), wherein a constant temperature oscillation mechanism (2), a metal bath mechanism (3), two water bath mechanisms (4), a liquid transfer mechanism and a transport mechanism are arranged in the workstation box (1); The pipetting mechanism is used to add each sample into the deep well plate (6); The constant temperature shaking mechanism (2) is used to shake and resuscitate the sample in the deep well plate (6) at a constant temperature; The metal bath mechanism (3) is used to keep the samples in the PCR plate at a constant temperature; The two water bath mechanisms (4) are used to perform cold bath treatment and heat shock treatment on the samples in the deep well plate (6) respectively; The transport mechanism is used to transport the deep well plate (6).

2. The automated cell transformation workstation according to claim 1, characterized in that: The water bath mechanism (4) comprises a water bath (5) arranged in the workstation housing (1), the water bath (5) being detachably connected to the deep-well plate (6), a liquid supply component being arranged in the workstation housing (1), the liquid supply component being connected to the water bath (5), a cooling and heating component being arranged on the water bath (5), and a temperature sensor (25) being fixedly connected in the water bath (5).

3. The automated cell transformation workstation according to claim 2, characterized in that: The cooling and heating component comprises a semiconductor cooling and heating plate (7) fixedly connected to the bottom of the water bath (5); a copper heat conducting pipe (8) is fixedly connected to the bottom of the water bath (5); the semiconductor cooling and heating plate (7) is in contact with the copper heat conducting pipe (8) and the outer bottom wall of the water bath (5).

4. The automated cell transformation workstation according to claim 3, characterized in that: A water-cooled copper bar is also provided at one end of the semiconductor refrigeration and heating plate (7) away from the water bath (5), and the water-cooled copper bar comprises a copper shell (9) fixedly connected to the semiconductor refrigeration and heating plate (7), a first circulation pipe (10) and a second circulation pipe (11) are fixedly connected to and connected to the copper shell (9), the first circulation pipe (10) is fixedly connected to and connected to a water cooling box (12), a water pump is fixedly connected inside the water cooling box (12), and a water outlet end of the water pump is fixedly connected to and connected to the second circulation pipe (11).

5. The automated cell transformation workstation according to claim 2, characterized in that: A plurality of spring sheets are fixedly connected to the outer wall of the water bath (5) in the circumferential direction, and the deep hole plate (6) is sandwiched between the plurality of spring sheets.

6. The automated cell transformation workstation according to claim 2, characterized in that: The liquid supply assembly comprises a liquid bottle (13) fixedly connected to the workstation housing (1); a liquid inlet is provided on the water bath (5); the liquid bottle (13) is fixedly connected to and communicated with a water inlet of a peristaltic pump (14) via a first liquid pipe; and a water outlet of the peristaltic pump (14) is fixedly connected to and communicated with the liquid inlet via a second liquid pipe.

7. The automated cell transformation workstation according to claim 2, characterized in that: The outer cover of the water bath (5) is provided with a heat insulation cover (15) with an opening at the top, and a heat insulation cover (16) is provided on the top of the heat insulation cover (15). The heat insulation cover (15) is fixedly connected to the workstation box (1), and the heat insulation cover (16) covers the top of the deep well plate (6).

8. The automated cell transformation workstation according to claim 2, characterized in that: A first support plate (17) is fixedly connected inside the workstation housing (1), and the water bath (5), the constant temperature oscillation mechanism (2), and the metal bath mechanism (3) are all fixedly connected to the first support plate (17); the liquid transfer mechanism comprises a motion component fixedly connected inside the workstation housing (1), an output end of the motion component is fixedly connected to an electric multi-channel liquid transfer gun (21), and the motion component drives the electric multi-channel liquid transfer gun (21) to move along the X-axis, Y-axis, and Z-axis directions.

9. The automated cell transformation workstation according to claim 1, characterized in that: An air filter (22) and an ultraviolet lamp (23) are fixedly connected to the inner top wall of the workstation box (1).

10. A working method for automated cell transformation, according to the automated cell transformation workstation according to any one of claims 1 to 9, characterized in that: The following steps are involved: Step 1: Pre-start The constant temperature oscillation mechanism (2), the metal bath mechanism (3) and the two water bath mechanisms (4) are started, and the preheating and precooling temperatures are set and made to reach the set values; and the workstation is disinfected at the same time; Step 2: Place the actual consumables in the designated location Place a 1000uL pipette tip, a 20uL pipette tip, a PCR plate containing a DNA sample, a deep-well plate (6) containing competent cells, a SOC culture medium, a heat-insulating cover (16), and a sponge pad at designated locations respectively; Step 3: Lysis of competent cells The transport mechanism transfers the heat-insulating cover (16) to the deep-well plate (6), and leaves it to stand for 15 minutes to allow the competent cells to slowly dissolve in the pre-cooled water bath mechanism (4); Step 4: Sample mixing After the standing is completed, the transport mechanism moves the heat-insulating cover (16) away from the deep-well plate (6) and returns it to its original position. The pipetting mechanism absorbs all DNA samples from the PCR plate and adds them to the deep-well plate (6). The samples are mixed with the competent cells in the deep-well plate (6) by in-situ aspiration and discharge. The heat-insulating cover (16) is then covered on the deep-well plate (6) by the transport mechanism and allowed to stand for 30 minutes to allow the DNA samples to be in contact with the competent cells and to be stable. Step 5: Heat shock treatment After the static state is completed, the transport mechanism quickly transports the deep-well plate (6) to another preheated water bath mechanism (4) for heat shock treatment; Step 6: Cold Bath Treatment After the heat shock is completed, the transport mechanism quickly transfers the deep well plate (6) back to the pre-cooled water bath mechanism (4) for cooling and standing; Step 7: Adding culture medium The transport mechanism removes the heat-insulating cover on the deep-well plate (6), and the pipetting mechanism draws a required amount of SOC culture medium from the liquid reservoir and adds it to the deep-well plate (6); Step 8: Constant temperature shaking culture The transport mechanism transfers the deep-well plate (6) to the constant temperature shaking mechanism (2), and then transports the heat-insulating cover back to cover the deep-well plate (6) again, and shakes the plate at a constant temperature for 1 hour to revive the transformed competent cells.

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