CN110835598B - Loading device, loading method and gene sequencing system - Google Patents
Loading device, loading method and gene sequencing system Download PDFInfo
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- CN110835598B CN110835598B CN201810936549.XA CN201810936549A CN110835598B CN 110835598 B CN110835598 B CN 110835598B CN 201810936549 A CN201810936549 A CN 201810936549A CN 110835598 B CN110835598 B CN 110835598B
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- C12Q1/6874—Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation
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Abstract
The invention provides a loading device for loading different liquid reactants on a sequencing chip to perform biochemical reaction, which comprises a cover plate and a base, wherein the bottom surface of the cover plate is provided with a sealing groove and a flow channel groove, the flow channel groove is arranged on the inner side of the sealing groove, the cover plate is abutted against the sequencing chip through a sealing piece embedded in the sealing groove to form a cavity, the flow channel groove is provided with a liquid inlet and a liquid outlet, the liquid inlet or the liquid outlet passes across the sealing groove through a corresponding flow channel in the cover plate and is communicated with an electromagnetic valve on the base, and the electromagnetic valve is used for controlling liquid adding and liquid discharging. The application of the device ensures that the sequencing chip does not need expensive packaging treatment, the reactant can be directly loaded on the bare sequencing chip, the cost is low, the operation is convenient, no bubble exists in the loading process, the liquid change is stable, and the rapid and large-flux automatic biological reaction is facilitated.
Description
Technical Field
The invention relates to the technical field of genes, in particular to a loading device, a loading method and a gene sequencing system using the device, wherein the loading device is used for loading a plurality of liquid reactants on a bare sequencing chip to perform biochemical reaction.
Background
This section is intended to provide a background or context to the embodiments of the invention that are recited in the claims. The description herein is not admitted to be prior art by inclusion in this section.
The gene sequencing chip (such as a silicon chip) needs to carry out biological reaction with a plurality of different reagents to fix or carry gene molecules or nucleic acid molecules on the surface of the chip, the liquid changing speed, liquid changing amount, liquid changing time, liquid changing temperature and liquid changing stability in the process of loading the reagents need to be strictly controlled, and the chip cannot be contacted with air in the process of changing the reagents, so that the chip loading reaction process needs to be carried out in a specially manufactured sealing device. Taking the loading of gene nanosphere molecules (DNB) as an example, the prior art scheme is a silicon chip closed DNB loading method, and the method is characterized in that each chip is specially processed and packaged in advance (firstly, a glass sheet is covered at a front fixed position of the chip, a runner is formed inside the chip by edge dispensing packaging, two ends of the runner are correspondingly provided with a liquid inlet and a liquid outlet on the chip, and the combined chip and glass sheet are fixed by a frame), the packaged chip is communicated with DNB or a reagent to the liquid inlet, negative pressure is generated by pumping liquid from an injector of the liquid outlet, a reactant is sucked into the runner from the liquid inlet and fully filled, and then biological reaction is carried out, and the reactant in the runner is continuously pumped and replaced until the reaction is complete.
According to the silicon wafer DNB loading method, the packaging processing of each chip is complex, the requirements on the precision and the tightness of a flow channel are extremely high, and the rejection rate of the chips during packaging is high; meanwhile, the auxiliary materials (glass sheets and frames) have large demand and high cost; moreover, each chip needs to be packaged, which is not beneficial to flexible use, especially the chip is difficult to remove once the air bubbles enter, and is not beneficial to rapid and large-flux automatic biological reaction after the glass sheet is packaged.
Disclosure of Invention
In view of the foregoing, there is a need for an improved loading device, loading method and gene sequencing system for loading several liquid reagents on a bare sequencing chip that can perform rapid, high throughput automated biological reactions.
The technical scheme provided by the invention is as follows: the utility model provides a loading device for load the liquid reactant and carry out biochemical reaction on sequencing chip, includes apron and base, set up seal groove and runner groove on the apron bottom surface, the runner groove is located the inboard of seal groove, the apron through the embedding the sealing member of seal groove contradict in on the sequencing chip and form the cavity, the runner groove is equipped with inlet and liquid outlet, the inlet or the liquid outlet passes through the inside corresponding runner of apron is strideed across the seal groove, and with solenoid valve intercommunication on the base, the solenoid valve is used for control liquid feeding and flowing back.
Further, the cavity is square.
Further, the runner groove is provided with a plurality of runner grooves, each runner groove is parallel to the inner side edge of the sealing piece, and the end part of each runner groove is provided with a plurality of liquid inlets and liquid outlets.
Specifically, the runner groove is provided with 2, is offered along relative sealing member inboard edge respectively, and its tip is equipped with 1 inlet, 3 liquid outlets.
Further, a second sealing element is arranged at the interface of the liquid inlet or the liquid outlet and the base.
Further, the second sealing element is embedded into the cover plate and abuts against the base.
Further, the electromagnetic valve comprises a corresponding number of liquid inlet valves, liquid outlet valves and an exhaust valve, wherein the liquid inlet valves are connected with the liquid inlet through pipelines to form liquid feeding flow paths, each liquid outlet valve is connected with a corresponding liquid outlet and a liquid outlet through pipelines to form liquid outlet flow paths, and the exhaust valve is connected with the liquid inlet, the liquid inlet valves, at least one liquid outlet valve and the liquid outlet through pipelines to form exhaust flow paths.
Specifically, the electromagnetic valve is provided with 5 electromagnetic valves, and the electromagnetic valve comprises 1 liquid inlet valve, 1 exhaust valve and 3 liquid discharge valves, wherein the liquid inlet valve is connected with a liquid inlet through a pipeline to form a liquid feeding flow path, each liquid discharge valve is connected with a corresponding liquid outlet and a liquid outlet through a pipeline to form a liquid discharge flow path, and the exhaust valve is connected with the liquid inlet, the liquid inlet valve, at least one liquid discharge valve and the liquid outlet through pipelines to form an exhaust flow path.
Further, the base is provided with a first heat conducting plate, a valve seat and a second heat conducting plate from top to bottom in sequence, the valve seat is of a hollow structure, the bottom of the first heat conducting plate is in contact with and fixed to the second heat conducting plate, the upper surface of the side edge of the first heat conducting plate is in contact with the cover plate and is fixed by a screw, and the lower surface of the first heat conducting plate is in contact with the valve seat.
Further, the top surface of the first heat-conducting plate is provided with a groove with one side communicated with the outside and used for clamping the sequencing chip, and after the sequencing chip is installed, the top surface of the sequencing chip is flush with the upper surface of the side edge of the first heat-conducting plate.
Further, the groove outlet of the first heat-conducting plate is used for extending the sequencing chip, and the extending part is provided with a clamping piece.
Further, the cover plate and the base are fixed by screws to realize the depth adjustment of the cavity, and the cover plate is provided with a positioning through hole.
The invention also provides a loading method for loading different liquid reactants on a sequencing chip for biochemical reaction, and the loading device is adopted for loading, which comprises the following steps:
when liquid is added for the first time, the exhaust valve is opened, liquid is sucked in, and air in the pipeline is removed;
opening the liquid inlet valve and the first liquid outlet valve, and closing the exhaust valve, wherein liquid flows to the first liquid outlet along the first flow channel groove;
when the liquid reaches the first liquid outlet, the first liquid discharge valve is closed, the second liquid discharge valve is opened, and the liquid flows to the second liquid outlet in parallel to the first flow channel groove;
when the liquid reaches the second liquid outlet, closing the second liquid discharge valve, opening the third liquid discharge valve, filling the second runner groove to the third liquid outlet by the liquid, and closing the liquid inlet valve and the third liquid discharge valve until the reaction is complete;
when the liquid is changed, the steps are repeated, the liquid in the pipeline is removed, and then the liquid in the cavity is gradually changed and then reacted.
Further, the end part of the first runner groove is provided with the liquid inlet and the first liquid outlet respectively, the second runner groove is parallel to the first runner groove, the end part of the second runner groove is provided with the second liquid outlet and the third liquid outlet respectively, and the liquid inlet is adjacent to the third liquid outlet.
The invention further provides a gene sequencing system, which is a bare sequencing chip soaking reaction testing system, and comprises a fluid system, a heating system and the loading device, wherein the fluid system is communicated with the infusion port, the opening and closing of the electromagnetic valve are controlled by an automatic program, and the heating system is contacted with the bottom of the second heat conducting plate to provide and control the temperature required by the reaction.
Compared with the prior art, the loading device is used for loading a liquid reactant on a sequencing chip to perform biochemical reaction, and comprises a cover plate and a base, wherein the bottom surface of the cover plate is provided with a sealing groove and a flow channel groove, the flow channel groove is arranged on the inner side of the sealing groove, the cover plate is abutted against the sequencing chip through a sealing piece embedded in the sealing groove to form a cavity, the flow channel groove is provided with a liquid inlet and a liquid outlet, the liquid inlet or the liquid outlet passes through the sealing groove through a corresponding flow channel in the cover plate, and is communicated with an electromagnetic valve on the base, and the electromagnetic valve is used for controlling liquid feeding and liquid discharging. The application of the device ensures that the sequencing chip does not need expensive packaging treatment, the reactant can be directly loaded on the bare sequencing chip, the cost is low, and the operation is convenient; the loading method is simple and efficient, has no bubble in the loading process, is stable in liquid change, and is beneficial to rapid and large-flux automatic biological reaction; the gene sequencing system can automatically load different reactants onto the bare sequencing chip, so that the bare sequencing chip carries gene molecules or nucleic acid molecules, the utilization rate of the sequencing chip is improved, and the reaction flux is high; the requirement of repeated verification test can be met in a single time, the test efficiency is high, and the test result is more accurate; meanwhile, the automatic loading program control is combined with the cavity design, so that the system error of multiple loads is small, and the test result is more reliable.
Drawings
The invention will be described in further detail with reference to the drawings and the detailed description.
Fig. 1 is a perspective view of a loading device according to an embodiment of the present invention.
FIG. 2 is an exploded view of the loading device of FIG. 1 from a perspective.
FIG. 3 is an exploded view of the loading device shown in FIG. 1 from another perspective.
Fig. 4 is a longitudinal section of the cover plate shown in fig. 1 at the liquid inlet.
Fig. 5 is a bottom view of the cover plate shown in fig. 1.
Reference numerals illustrate:
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100 |
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10 |
| |
11 |
| |
12 |
| Positioning through |
13 |
| Threaded through |
14 |
| |
15 |
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16 |
| Liquid inlet | 161 |
| A first liquid outlet | 162 |
| A second |
163 |
| Third |
164 |
| First |
20 |
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30 |
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40 |
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41 |
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42 |
| First |
43 |
| Second |
44 |
| Third |
45 |
| Infusion port | 1 |
| Waste |
2 |
| |
50 |
| Clamping |
51 |
| Second |
60 |
The following detailed description will further illustrate embodiments of the invention in conjunction with the above-described drawings.
Detailed Description
In order that the above-recited objects, features and advantages of embodiments of the present invention can be more clearly understood, a more particular description of the invention will be rendered by reference to the appended drawings and appended detailed description. In addition, features in the embodiments of the present application may be combined with each other without conflict.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of embodiments of the invention, and the described embodiments are merely some, rather than all, of the embodiments of the invention. All other embodiments, based on the embodiments of the invention, which are obtained by a person of ordinary skill in the art without making any inventive effort, are within the scope of the embodiments of the invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the invention belong. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The gene sequencing system is used for detecting the base pair sequencing rule on the gene molecules or the nucleic acid molecules and analyzing and predicting diseases and risks. The system loads different liquid reactants dissolved with gene molecules or nucleic acid molecules on a sequencing chip for full reaction, so that the surface of the sequencing chip carries a plurality of gene molecules or nucleic acid molecule main chains according to a certain rule, and then base pairing, fluorescent marking and photographing are carried out on groups on each main chain one by one, so that a complete sequence is obtained. Wherein, the loading of different liquid reactants with dissolved gene molecules or nucleic acid molecules onto the sequencing chip is the first key step, determining the sequencing throughput and sequencing quality in the later stage.
Because the sequencing chip 50 needs to perform biological reaction with a plurality of different reagents, the existing pre-packaging chip loading method has the disadvantages of high packaging processing cost, complexity, high rejection rate, low utilization rate of the sequencing chip 50, incapability of stopping air bubbles from entering, influence on the progress of biochemical reaction (the sequencing chip 50 cannot contact air in the reaction), and adverse effect on rapid and large-flux automatic biological reaction, so the invention designs the double-channel cover plate type loading device 100.
The overall structure of the loading device 100 of the present invention will be described in detail with reference to fig. 1 to 5.
The device 100 is used for loading liquid reactants on a sequencing chip to perform biochemical reaction, and the application of the device enables the sequencing chip 50 to be free from expensive packaging treatment, and the reactants can be directly loaded on the bare sequencing chip 50, so that the device is low in cost and convenient to operate.
Fig. 1 shows a main structure of the device 100, mainly comprising:
the cover plate 10 is covered on the sequencing chip 50 and forms a sealing structure with the sequencing chip 50;
the sequencing chip 50 is arranged between the cover plate 10 and the base, and a clamping piece 51 is arranged outside the sequencing chip, so that the sequencing chip is convenient to take;
a base, which supports the sequencing chip 50 from the bottom, and provides a heat source and mounts the solenoid valve 40.
The base further comprises a first heat-conducting plate 20 and a valve seat 30, electromagnetic valves 40 are arranged outside the left side edge and the right side edge of the valve seat 30, and an infusion port 1 and a waste liquid port 2 are arranged on the front side edge.
To describe the seal structure in detail, the internal components of the overall structure shown in fig. 1 are longitudinally exploded as shown in fig. 2 and 3.
In the present embodiment, the loading device 100 is provided with a cover plate 10, a sealing member 11, a second sealing member 12, a sequencing chip 50 (the outside of which is provided with a clamping member 51), a first heat-conducting plate 20, a valve seat 30 (mounted with a solenoid valve 40), and a second heat-conducting plate 60 in this order from the top to the bottom.
Referring to fig. 5, a sealing groove 15 is formed on the bottom surface of the cover plate 10. In this embodiment, the sealing groove 15 is a square groove on the bottom surface of the cover plate 10, the groove is in an inverted convex shape (as shown in fig. 4), the corresponding sealing element 11 is also in an inverted convex shape, the top of the sealing element 11 can be completely embedded into the sealing groove 15, and the bottom of the sealing element is abutted against the sequencing chip 50, so that a square cavity is formed among the cover plate 10, the sequencing chip 50 and the sealing element 11, the thickness of the cavity (i.e., the gap between the cover plate 10 and the sequencing chip 50) is usually controlled to be at or below a micron level, the loading effect is good (the consumption of reagents is less, the cost is reduced, and the loading efficiency is high); when the thickness is larger, the loading function can be realized, but the loading effect is not ideal.
It is to be understood that the shapes of the seal groove 15 and the seal 11 are not limited to the present embodiment; in other embodiments, the shape of the cavity is not limited to square, but may be circular or other.
As shown in fig. 5, a flow channel groove 16 is further provided on the bottom surface of the cover plate 10, and the flow channel groove 16 is provided inside the sealing groove 15. In this embodiment, the number of the flow channel grooves 16 is 2, the flow channel grooves 16 are parallel to one side of the sealing groove 15, the 2 flow channel grooves 16 are spaced apart from the inner side of the adjacent sealing groove 15, the sealing groove 15 and the flow channel grooves 16 are not in communication, and the 2 flow channel grooves 16 correspond to the liquid inlet direction and the liquid outlet direction, respectively. The function of the flow channel groove 16 is to effectively remove gas, prevent the sequencing chip 50 from contacting air during liquid exchange, and guide the liquid to flow.
It is understood that the number of the flow channel grooves 16 is not limited to 2, and may be one, or more than 2; in other embodiments, the flow channel groove 16 is not necessarily parallel to the seal groove 15, and does not necessarily correspond to the liquid inlet direction and the liquid outlet direction.
As further shown in fig. 5, the runner duct 16 is provided with a liquid inlet 161 and a plurality of liquid outlets (162, 163, 164), and as shown in fig. 4, the liquid inlet 161 or the liquid outlets (162, 163, 164) cross the seal groove 15 through corresponding runners inside the cover plate, and are communicated with the outside. The liquid inlet 161 or the interface between the liquid outlet (162, 163, 164) and the outside is provided with a second sealing member 12, and the second sealing member 12 is embedded into the cover plate 10. In this embodiment, the end of the runner groove 16 is provided with 1 liquid inlet 161 and 3 liquid outlets (162, 163, 164), which are respectively located at 4 inner corners of the seal groove 15. Correspondingly, the second sealing elements 12 are provided with 4, also inverted-convex. As can be seen in connection with fig. 2, the ends of the 4 second seals 12 abut against the valve seat 30, such that the inlet 161 or outlet is in communication with the solenoid valve 40 carried on the valve seat 30.
It is understood that the number of the liquid outlets (162, 163, 164) is not limited to the present embodiment; in other embodiments, the positions and the number of the liquid inlets 161 are not limited to the present embodiment.
The top edge of the cover plate 10 shown in fig. 2 is provided with a threaded through hole 14 penetrating through, and corresponds to the hole on the first heat conducting plate 20, and the threaded through hole is used for locking the two threads, so that the regulation and control of the depth of the cavity are realized.
It will be appreciated that the cover plate 10 and the first heat-conducting plate 20 are not necessarily screwed and locked, but may be clamped at the top and bottom, or in other manners, only needs to satisfy the fastening function and the regulation function.
As shown in fig. 2 and 3, the valve seat 30 is a hollow structure. In this embodiment, the electromagnetic valves 40 are provided with 5 liquid inlet valves 41, 1 air outlet valve 42 and 3 liquid outlet valves, wherein the liquid inlet valves 41 are connected with the liquid inlet 1 and the liquid inlet 161 by pipelines to form a liquid feeding flow path, each liquid outlet valve is connected with a corresponding liquid outlet and the liquid outlet 2 by pipelines to form a liquid outlet flow path, and the air outlet valves 42 are connected with the liquid inlet 1, the liquid inlet valves 41, at least one liquid outlet valve and the liquid outlet 2 by pipelines to form an air outlet flow path. In the specific embodiment, the liquid inlet valve 41, the air outlet valve 42 and the first liquid outlet valve 43 are sequentially arranged outside the left side surface of the valve seat 30, wherein the liquid inlet valve 41 is nearest to the liquid inlet 1 and is communicated with the liquid inlet, the second liquid outlet valve 44 and the third liquid outlet valve 45 are arranged outside the right side surface of the valve seat 30, and the third liquid outlet valve 45 is nearest to the liquid outlet 2. The liquid inlet 161, the liquid inlet valve 41 and the liquid delivery port 1 on the first runner groove 16 are communicated, the first liquid outlet 162, the first liquid outlet valve 43 and the liquid outlet 2 on the first runner groove 16 are communicated, the second liquid outlet 163, the second liquid outlet valve 44 and the liquid outlet 2 on the second runner groove 16 are communicated, the third liquid outlet 164, the third liquid outlet valve 45 and the liquid outlet 2 on the second runner groove 16 are communicated, one port of the exhaust valve 42 is communicated with the liquid inlet valve 41, and the other port of the exhaust valve 42 is simultaneously communicated with 3 liquid discharge flow paths, so that the exhaust valve 42 can remove gas in a pipeline and liquid (various reactants) to be replaced. The exhaust valve 42 is a bypass in nature and can discharge liquid that is not allowed to flow into the liquid inlet 161 directly to the liquid outlet 2. For example, when the liquid flowing in through the infusion port 1 contains bubbles or the liquid is a pipe cleaning liquid, the liquid can be directly discharged through the exhaust valve 42 without flowing into the liquid inlet 161.
It will be appreciated that the number and positions of the liquid inlet 161 and the liquid outlet are not limited, and accordingly, the number and functions of the electromagnetic valve 40 are not limited to the above configuration, and the operations of exhausting, charging and discharging can be satisfied.
The sequencing chip 50 is placed on the first heat-conducting plate 20, and the top surface of the sequencing chip is surrounded by the cover plate 10 and the sealing member 11 to form a cavity. In this embodiment, the sequencing chip 50 in the cavity sealing area is square, the edge of the sequencing chip 50 is clamped on the first heat-conducting plate 20, and the top surface of the sequencing chip 50 is flush with the upper surface of the side edge of the first heat-conducting plate 20. The area of the sequencing chip 50 extending out of the first heat-conducting plate 20 is in a sector shape, and the sector part is provided with a clamping piece 51, specifically, the clamping piece 51 is arranged above the sequencing chip 50 and is fixed in an adhesion manner, so that the sequencing chip is convenient to take and does not pollute the surface of the sequencing chip 50 carrying the gene molecules or the nucleic acid molecules.
It is to be understood that the shape of the sequencing chip 50 is not limited to the present embodiment, and may be as high as possible, and may be ensured to be clamped.
The bottom of the first heat-conducting plate 20 is in contact with the second heat-conducting plate 60 and is screwed upwards from the bottom of the second heat-conducting plate 60, the upper side surface of the first heat-conducting plate 20 is in contact with the cover plate 10 and is screwed, and the lower side surface of the first heat-conducting plate is in contact with and is clamped with the valve seat 30. The cover plate 10 is provided with a positioning through hole 13 which is positioned with the first heat conducting plate 20 by a bolt. The shapes of the first heat-conducting plate 20 and the second heat-conducting plate 60 are set according to actual needs, and will not be described herein.
The priming process of the loading device 100 of the present invention is described in further detail below in conjunction with fig. 5.
Step 1: when the liquid is added for the first time, the exhaust valve 42 is opened, liquid is sucked in, and the air in the pipeline is removed;
step 2: opening the liquid inlet valve 41 and the first liquid outlet valve 43, and closing the air outlet valve 42, so that the liquid flows to the first liquid outlet 162 along the first flow channel 16;
step 3: when the liquid reaches the first liquid outlet 162, the first liquid discharging valve 43 is closed, the second liquid discharging valve 44 is opened, and the liquid flows to the second liquid outlet 163 parallel to the first flow channel 16;
step 4: when the liquid reaches the second liquid outlet 163, the second liquid discharge valve 44 is closed, the third liquid discharge valve 45 is opened, the liquid fills the second flow channel 16 to the third liquid outlet 164, and the liquid inlet valve 41 and the third liquid discharge valve 45 are closed for complete reaction.
The liquid change process of the loading device 100 of the present invention is further specifically described below with reference to fig. 5.
Step 1: opening the exhaust valve 42 to suck new liquid and replace the reacted liquid (waste liquid) in the pipeline;
step 2: opening the liquid inlet valve 41 and the first liquid discharge valve 43, and closing the air outlet valve 42, so that the liquid flows to the first liquid outlet 162 along the first flow channel 16, and the waste liquid is pumped out of the first liquid outlet 162 or extruded to the liquid discharge flow path until being discharged from the waste liquid outlet 2;
step 3: when the liquid reaches the first liquid outlet 162, the first liquid discharge valve 43 is closed, the second liquid discharge valve 44 is opened, the liquid flows to the second liquid outlet 163 parallel to the first flow channel 16, and meanwhile, the waste liquid is pumped out of the second liquid outlet 163 or extruded into the liquid discharge flow channel until being discharged from the waste liquid outlet 2; the second drain valve 44 is opened for a longer period than the first filling, mainly because the viscosity of the liquid itself causes a lower flow rate than the evacuation rate of the gas.
Step 4: when the liquid reaches the second liquid outlet 163, the second liquid discharge valve 44 is closed, the third liquid discharge valve 45 is opened, the liquid fills the second flow channel 16 to the third liquid outlet 164, and the waste liquid is pumped out of the third liquid outlet 164 or extruded to the liquid discharge flow path until the waste liquid is discharged from the waste liquid outlet 2, and the liquid inlet valve 41 and the third liquid discharge valve 45 are closed for complete reaction.
In conclusion, the loading method using the device is simple, efficient, bubble-free in the loading process, stable in liquid change and beneficial to rapid and large-flux automatic biological reaction.
The gene sequencing system using the loading device 100 provided by the invention also comprises a fluid system and a heating system.
Wherein:
the fluid system is communicated with the infusion port 1, and the opening and closing of the electromagnetic valve 40 are controlled by an automatic program;
the heating system is in contact with the bottom of the second heat-conducting plate 60 to provide and control the temperature required for the reaction.
The system can automatically load various liquid reactants onto the bare sequencing chip 50, the liquid exchange process is efficient and stable, no bubbles exist, the utilization rate of the sequencing chip 50 is improved, and the reaction flux is high; the requirement of repeated verification test can be met in a single time, the test efficiency is high, and the test result is more accurate; meanwhile, the automatic loading program control is combined with the design of the sealed cavity, so that the systematic error of multiple loading is small, and the test result is more reliable.
The foregoing embodiments are merely for illustrating the technical solution of the embodiment of the present invention, but not for limiting the same, although the embodiment of the present invention has been described in detail with reference to the foregoing preferred embodiments, it will be understood by those skilled in the art that modifications and equivalent substitutions may be made to the technical solution of the embodiment of the present invention without departing from the spirit and scope of the technical solution of the embodiment of the present invention.
Claims (8)
1. A loading device for loading a liquid reactant on a sequencing chip for biochemical reaction, characterized in that: the sequencing chip sequencing device comprises a cover plate and a base, wherein a sealing groove and a flow channel groove are formed in the bottom surface of the cover plate, the flow channel groove is formed in the inner side of the sealing groove, the cover plate is abutted against the sequencing chip through a sealing piece embedded in the sealing groove and forms a cavity, the flow channel groove is provided with a liquid inlet and a liquid outlet, the liquid inlet or the liquid outlet spans the sealing groove through a corresponding flow channel in the cover plate and is communicated with an electromagnetic valve on the base, and the electromagnetic valve is used for controlling liquid adding and liquid discharging;
the cavity is square, the flow channel grooves are provided with a plurality of flow channel grooves, each flow channel groove is arranged parallel to the edge of the inner side of the sealing element, the end part of each flow channel groove is provided with a plurality of liquid inlets and liquid outlets, and a second sealing element is arranged at the interface of the liquid inlet or the liquid outlet and the base;
the electromagnetic valve comprises a corresponding number of liquid inlet valves, liquid outlet valves and an exhaust valve, wherein the liquid inlet valves are connected with the liquid inlet through pipelines to form liquid adding flow paths, each liquid outlet valve is connected with a corresponding liquid outlet and a liquid outlet through pipelines to form liquid discharging flow paths, and the exhaust valve is connected with the liquid inlet, the liquid inlet valves, at least one liquid outlet valve and the liquid outlet through pipelines to form exhaust flow paths.
2. The loading device of claim 1, wherein: the base top-down is equipped with first heat-conducting plate, disk seat and second heat-conducting plate in proper order, the disk seat is hollow structure, the bottom of first heat-conducting plate with the second heat-conducting plate contacts and fixes, the side upper surface of first heat-conducting plate with the apron contacts and the screw is fixed, and the lower surface with the disk seat contacts.
3. The loading device of claim 2, wherein: the top surface of the first heat-conducting plate is provided with a groove with one side communicated with the outside and used for clamping the sequencing chip, and after the sequencing chip is installed, the top surface of the sequencing chip is flush with the upper surface of the side edge of the first heat-conducting plate.
4. The loading device of claim 2, wherein: the groove outlet of the first heat-conducting plate is used for extending out of the sequencing chip.
5. The loading device of claim 1, wherein: the cover plate and the base are fixed by screws to realize the depth adjustment of the cavity, and the cover plate is provided with a positioning through hole.
6. A loading method for loading a liquid reagent on a sequencing chip for biochemical reaction, characterized in that the loading is performed by using the loading device according to any one of claims 1 to 5, comprising the steps of:
opening the exhaust valve, sucking liquid and removing pipeline air;
opening the liquid inlet valve and the first liquid outlet valve, and closing the exhaust valve, wherein liquid flows to the first liquid outlet along the first flow channel groove;
when the liquid reaches the first liquid outlet, the first liquid discharge valve is closed, the second liquid discharge valve is opened, and the liquid flows to the second liquid outlet in parallel to the first flow channel groove;
when the liquid reaches the second liquid outlet, the second liquid discharge valve is closed, the third liquid discharge valve is opened, the liquid fills the second runner groove to the third liquid outlet, the liquid inlet valve and the third liquid discharge valve are closed until the reaction is complete, and the waste liquid is discharged.
7. The loading method according to claim 6, wherein: the end part of the first runner groove is respectively provided with the liquid inlet and the first liquid outlet, the second runner groove is parallel to the first runner groove, the end part of the second runner groove is respectively provided with the second liquid outlet and the third liquid outlet, and the liquid inlet is adjacent to the third liquid outlet.
8. A gene sequencing system, characterized in that: comprising a fluid system in communication with the infusion port and automatically programmed to open and close the solenoid valve, a heating system providing and controlling the temperature required for the reaction, and a loading device according to any one of claims 1 to 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810936549.XA CN110835598B (en) | 2018-08-16 | 2018-08-16 | Loading device, loading method and gene sequencing system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810936549.XA CN110835598B (en) | 2018-08-16 | 2018-08-16 | Loading device, loading method and gene sequencing system |
Publications (2)
| Publication Number | Publication Date |
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| CN110835598A CN110835598A (en) | 2020-02-25 |
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