CN220887449U - Digital PCR pore plate - Google Patents
Digital PCR pore plate Download PDFInfo
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- CN220887449U CN220887449U CN202322049722.3U CN202322049722U CN220887449U CN 220887449 U CN220887449 U CN 220887449U CN 202322049722 U CN202322049722 U CN 202322049722U CN 220887449 U CN220887449 U CN 220887449U
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- digital pcr
- open container
- well plate
- flat bottom
- pcr well
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- 238000007847 digital PCR Methods 0.000 title claims abstract description 43
- 239000011148 porous material Substances 0.000 title claims abstract description 21
- 230000003014 reinforcing effect Effects 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 19
- 239000002699 waste material Substances 0.000 claims description 17
- 229920001296 polysiloxane Polymers 0.000 claims description 15
- 239000003292 glue Substances 0.000 claims description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 239000000499 gel Substances 0.000 claims description 2
- 238000001746 injection moulding Methods 0.000 abstract description 14
- 238000001514 detection method Methods 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 238000003384 imaging method Methods 0.000 abstract description 3
- 238000010438 heat treatment Methods 0.000 description 8
- 238000003754 machining Methods 0.000 description 7
- 150000007523 nucleic acids Chemical class 0.000 description 6
- 102000039446 nucleic acids Human genes 0.000 description 6
- 108020004707 nucleic acids Proteins 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- 229910000838 Al alloy Inorganic materials 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010222 PCR analysis Methods 0.000 description 1
- 230000004308 accommodation Effects 0.000 description 1
- 238000004026 adhesive bonding Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000012632 fluorescent imaging Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000004381 surface treatment Methods 0.000 description 1
- 238000010301 surface-oxidation reaction Methods 0.000 description 1
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- Optical Measuring Cells (AREA)
Abstract
The utility model provides a digital PCR pore plate which relates to the technical field of biomedical detection, comprising an injection-molded open container, wherein the open container is provided with a bottom plate part and a surrounding part, and the surrounding part is arranged around the edge of the bottom plate part; a plurality of flat bottom sample tanks which are sequentially arranged are arranged on the bottom plate part; the roughness of the inner bottom surface of the flat bottom sample cell is 0.1-0.2 mu m. The roughness of the outer bottom surface of the open container is 0.02-0.06 mu m. According to the utility model, the digital PCR pore plate is changed into injection molding production, and the roughness of the observation surface is designed due to the influence of the roughness of the observation surface on imaging detection, so that the cost is greatly reduced, and the digital PCR pore plate formed by injection molding provides excellent observation conditions.
Description
Technical Field
The utility model relates to the technical field of biomedical detection, in particular to a digital PCR pore plate.
Background
Microdroplet digital PCR (droplet digital polymerase chain reaction, ddPCR) is a third generation PCR technique, a method of absolute quantification of nucleic acid analysis. The ddPCR works on the principle that the reaction solution is subjected to microdroplet treatment before PCR amplification, namely the reaction solution containing the nucleic acid template is dispersed into tens of thousands of nano-scaled micro-droplets, each micro-droplet does not contain or contains one to a plurality of target molecules of the nucleic acid to be detected, and each micro-droplet is used as an independent PCR reaction unit. After PCR amplification, the micro-droplets containing the target molecules of the nucleic acids to be detected generate fluorescence signals, and the micro-droplets not containing the target molecules of the nucleic acids to be detected do not generate fluorescence signals. And finally, calculating the initial concentration or copy number of the target molecules of the nucleic acid to be detected according to the poisson distribution principle and the proportion of positive micro-droplets.
At present, oily liquid can be injected into a digital PCR pore plate, a sample injection needle is adopted to vibrate under the liquid level of the pore plate to generate micro-droplets, a heating device is adopted to perform program temperature control on the pore plate filled with the micro-droplets to realize PCR amplification, and finally excitation light is used to irradiate the micro-droplets paved inside the pore plate to perform fluorescence imaging detection. The existing digital PCR pore plate adopts an aluminum alloy machine part and surface oxidation black treatment, so that the surface of the pore plate can meet the requirements of micro-droplet fluorescent imaging detection. However, the cost of the oxidation treatment of the aluminum alloy machining parts is high, the aluminum alloy machining parts are not beneficial to large-batch use, the environment of the aluminum alloy machining parts and the surface treatment is easy to cause greasy dirt and dust pollution of the pore plate, the test result is affected, and if the post-cleaning and drying treatment is carried out, the cost is further increased.
Disclosure of utility model
In view of the above, the present application provides a digital PCR well plate for solving at least one of the above technical problems, and provides a digital PCR well plate with low cost and convenient imaging detection.
The utility model provides a digital PCR pore plate, which comprises an injection-molded open container,
The open container has a bottom plate portion and a surrounding portion provided around an edge of the bottom plate portion; a plurality of flat bottom sample tanks which are sequentially arranged are arranged on the bottom plate part;
The roughness of the inner bottom surface of the flat bottom sample cell is 0.1-0.2 mu m.
In a preferred embodiment, the roughness of the outer bottom surface of the open container is 0.02-0.06 μm.
In a preferred embodiment, the bottom plate portion of the open container is provided with edge ribs and inner hole ribs; the edge reinforcing ribs are arranged around the bottom plate part, the inner hole reinforcing ribs are arranged inside the edge reinforcing ribs in a crisscross mode, and the inner space surrounded by the edge reinforcing ribs is divided into a plurality of flat bottom sample tanks.
In a preferred embodiment, the flat bottom sample cell is rectangular in structure, and the inner corners of the side walls of the flat bottom sample cell are circular arc structures.
In a preferred embodiment, two strip-shaped waste liquid tanks are oppositely arranged on the edge reinforcing ribs in the open container.
In a preferred embodiment, the inner edge reinforcing rib of the open container is further provided with a plurality of linearly arranged glue reducing holes; the gel reducing hole is positioned between two adjacent flat bottom sample tanks.
In a preferred embodiment, the digital PCR well plate further comprises a thermally conductive silicone sheet disposed at the outer bottom of the open container.
In a preferred embodiment, the thermally conductive silicone sheet has a hardness of 30 to 50; the thickness of the heat conduction silica gel sheet is 0.5mm-1.5mm.
In a preferred embodiment, the surrounding portion is provided obliquely to the bottom plate portion.
In a preferred embodiment, the top of the skirt extends outwardly to form a rim which is integrally formed with the open container.
In a preferred embodiment, the rim is provided with an identification notch.
Compared with the prior art, the utility model changes the digital PCR pore plate from metal machining to injection molding production. The digital PCR pore plate produced by injection molding can greatly reduce the production cost, design the roughness of the inner bottom surface of the flat bottom sample cell and improve the accuracy of observation.
The above technical features can be combined in various technically feasible ways to create new embodiments as long as the object of the utility model is achieved.
Drawings
FIG. 1 is a first perspective view of a digital PCR well plate according to an embodiment of the present utility model;
FIG. 2 is a front view of a digital PCR well plate according to an embodiment of the present utility model;
FIG. 3 is a second perspective view of a digital PCR well plate according to an embodiment of the present utility model;
FIG. 4 is a third perspective view of a digital PCR well plate according to an embodiment of the present utility model;
fig. 5 is a schematic diagram of a multi-segment curve structure according to an embodiment of the present utility model.
Wherein, the reference numerals are as follows:
1. A bottom plate portion; 2. a surrounding portion; 3. a flat bottom sample cell; 4. edge reinforcing ribs; 5. inner hole reinforcing ribs; 6. a waste liquid tank; 7. a glue reducing hole; 8. a thermally conductive silicone sheet; 9. an edge portion; 10. identifying a gap; 11-heating the base; 12. a multi-section curve structure; 121. a straight line portion; 122. an arc section.
Detailed Description
The following description of the embodiments of the present utility model will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the utility model. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
It is to be noted that unless otherwise defined, technical or scientific terms used herein should be taken in a general sense as understood by one of ordinary skill in the art to which the present utility model belongs. The terms "first," "second," and the like, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. In the description of the present utility model, the terms "upper," "lower," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like refer to an orientation or positional relationship based on that shown in the drawings, merely for convenience of description and simplicity of description, rather than to indicate or imply that the apparatus or element in question must have a specific orientation, be constructed and operated in a specific orientation, and that when the absolute position of the object being described is changed, the relative positional relationship may also be changed accordingly, and thus should not be construed as limiting the present utility model.
The technology not mentioned in the present utility model can be realized by the prior art.
As shown in fig. 1 to 4, the digital PCR well plate includes an injection-molded open container having a bottom plate portion 1 and a surrounding portion 2, and the bottom plate portion 1 may be of a rectangular structure as a whole and subjected to chamfering treatment at corner positions of the rectangular structure. The surrounding part 2 surrounds the edge of the bottom plate part 1 and is a side wall of the open container and is used for being matched with the bottom plate part 1 to form a complete open container. The bottom plate part 1 is provided with a plurality of flat bottom sample cells 3 arranged in sequence, and the flat bottom sample cells 3 are used for containing micro droplets. The digital PCR pore plate comprises an injection-molded open container, so that the production cost can be greatly reduced, and the injection molding can adopt a purification workshop for production and sealing, so that the test result can be prevented from being influenced by dust pollution and oil pollution of machining, and the further increase of the cost and the process risk caused by the post cleaning and drying treatment can be avoided.
Furthermore, the open container is black, so that the contrast ratio of the fluorescent signal of the positive micro-droplet to the background color can be increased, and the fluorescent signal can be conveniently observed. And the raw materials of the open container adopt a grain extraction process, so that the color of the finally formed open container can be consistent, and the influence on the test result caused by batch difference of the color depth of the open container is avoided.
Further, the dimensions and depth of each flat bottom sample cell 3 are the same, and the dimensions include the length, width, height, and curvature of the edges of the flat bottom sample cell 3. The roughness of the inner bottom surface of the flat bottom sample cell is 0.1-0.2 μm. If the smoothness of the inner bottom surface of the flat bottom sample cell 3 used as a background is too high, the light emitted by the excitation light forms reflected light on the inner bottom surface of the flat bottom sample cell 3, so that interpretation is affected. At the same time, the inner bottom surface of the flat bottom sample cell 3 is required to be uniform, and if too rough, the quality of observation is affected. Thus, considering the above two effects, the roughness of the inner bottom surface of the flat bottom sample cell 3 is set to 0.1 μm to 0.2 μm, and a balance is achieved between avoiding reflection of light and meeting the requirements of the flat bottom sample cell 3.
Further, the roughness of the inner bottom surface of the flat bottom sample cell 3 was one of 0.1 μm, 0.11 μm, 0.12 μm, 0.13 μm, 0.14 μm, 0.15 μm, 0.16 μm, 0.17 μm, 0.18 μm, 0.19 μm, 0.2 μm.
In a more preferred embodiment, the roughness of the outer bottom surface of the open container is 0.02 μm-0.06 μm. It will be appreciated that the microdroplet contained within the open container needs to be heated to conform to the conditions of the imaging assay. The entire open container can be heated by the heating base 11 to achieve indirect heating of the microdroplets. The machined metal digital PCR pore plate has high heat conductivity coefficient, but the roughness after machining is generally 0.8 mu m, the polishing treatment is carried out again, the cost is higher, the roughness is only 0.1 mu m under the ideal machining state, the contact area between the open container and the heating device is reduced due to the overhigh roughness, the heat transfer is not facilitated, and the temperature rise is still slower. In this embodiment, the roughness of the outer bottom surface of the open container is set to 0.02 μm-0.06 μm, so that the contact area between the digital PCR orifice plate formed by injection molding and the heating base 11 can be increased, and the heat transfer efficiency can be further improved.
Further, the roughness of the open container outer bottom surface is one of 0.021μm、0.022μm、0.023μm、0.024μm、0.025μm、0.026μm、0.027μm、0.028μm、0.029μm、0.03μm、0.031μm、0.032μm、0.033μm、0.034μm、0.035μm、0.036μm、0.037μm、0.038μm、0.039μm、0.04μm、0.041μm、0.042μm、0.043μm、0.044μm、0.045μm、0.046μm、0.047μm、0.048μm、0.049μm、0.05μm、0.051μm、0.052μm、0.053μm、0.054μm、0.055μm、0.056μm、0.057μm、0.058μm、0.059μm、0.06.
In a more preferred embodiment, edge ribs 4 and inner hole ribs 5 are provided on the bottom plate portion 1 in the open container. The edge reinforcing ribs 4 are located around the bottom plate portion 1. The inner hole reinforcing ribs 5 are arranged in the edge reinforcing ribs 4 in a crisscross manner, the inner space formed by the edge reinforcing ribs 4 is divided, flat bottom sample cells 3 are formed, and the flat bottom sample cells 3 are distributed on the bottom plate part 1 in an array manner.
In a more preferred embodiment, the flat bottom sample cell 3 is configured as a rectangular structure in order to conform to the injection molding process. The inner corners of the side walls of the flat bottom sample cell 3 of the rectangular structure are set to be circular arc structures.
In a more preferred embodiment, two strip-shaped waste liquid tanks 6 are oppositely arranged on the edge reinforcing ribs 4 in the open container, so that a sample adding needle can conveniently drop micro-droplets into the waste liquid tanks 6 firstly and then move into the flat-bottom sample tank 3 to add the micro-droplets in a fixed volume, and the accuracy of the quantity of the micro-droplets added into the flat-bottom sample tank 3 can be ensured due to the fact that the air at the head part of the sample adding needle is discharged in advance, and further the accuracy of digital PCR detection is improved. The waste liquid tank 6 is provided in a strip-like structure and is disposed relatively parallel to the edge reinforcing ribs 4 to increase the accommodating space of the waste liquid tank. Because the open container is formed by injection molding, if the local colloid volume is too large, the open container will shrink and deform after injection molding is completed, reducing the aesthetic appearance of the open container, and also causing deviation of the product size. In the embodiment, the waste liquid tank 6 is arranged to be of a downward concave structure, so that the glue consumption of the open container can be reduced during injection molding, and shrinkage and deformation after injection molding caused by too large local glue volume are avoided.
In order to further increase the accommodation space of the strip-shaped waste liquid tank 6, two waste liquid tanks 6 arranged opposite to each other on the edge reinforcing rib 4 are designed. The side of the strip-shaped waste liquid tank 6, which is close to the surrounding part 2, is of a linear structure, and the side of the strip-shaped waste liquid tank 6, which is close to the flat bottom sample tank 3, is provided with a multi-section curve structure 12; each multi-section curve structure 12 is arranged corresponding to the flat bottom sample cell 3. The multi-section curve structure 3 comprises a straight line part 121 and arc line parts 122 positioned at two ends of the straight line part 121, wherein the arc line parts 122 are parallel to the arc-shaped structures of the corresponding flat-bottom sample cells 3, and the straight line part 121 is parallel to the side edges of the flat-bottom sample cells 3. By designing the side edge of the waste liquid tank 6, the thickness of the reinforcing rib between the waste liquid tank 6 and the flat bottom sample tank 3 is consistent. By adopting the structure, the accommodating space of the waste liquid tank 6 can be increased, and shrinkage and deformation after injection molding caused by too large local glue volume can be further avoided.
In a more preferred embodiment, a waste liquid tank 6 is provided on each of opposite sides of the edge bead 4, and glue reducing holes 7 are provided on the other opposite sides of the edge bead 4. The glue reducing holes 7 are arranged linearly on one side of the edge reinforcing ribs 4 and are distributed between two adjacent flat bottom sample cells 3. The glue reducing hole 7 and the waste liquid tank 6 jointly realize glue reduction of the open container in the injection molding process.
In a more preferred embodiment, to speed up the detection process and further reduce the heating time, the digital PCR well plate further comprises a thermally conductive silicone sheet 8, one thermally conductive silicone sheet 8 being configured for the open container. As shown in fig. 2, the thermally conductive silicone sheet 8 may be adhered to the bottom of the open container by means of gluing. The heat conductive silicone sheet 8 can also be fixed to the open container bottom by means of two-shot molding.
Further, in order to further reduce the heating time, the heat conductive silicone sheet 8 may be designed. Since the harder the thermally conductive silicone sheet 8 is, the smaller the contact area with the bottom of the open container is, the smaller the volume of the gap between the open container and the thermally conductive silicone sheet 8 is filled, thereby affecting heat conduction. However, the thermal silicon sheet 8 is too soft, so that the thermal silicon sheet 8 deforms in the use process and cannot be adhered to the open container. Therefore, the hardness of the heat-conducting silica gel sheet 8 is designed to be between 30 and 50, and the unit is Shore hardness. Since the thickness of the thermally conductive silicone sheet 8 also affects the heat transfer efficiency, the thickness of the thermally conductive silicone sheet 8 is designed. The thickness of the heat conductive silicone sheet 8 may be any thickness between 0.5mm and 1.5mm, and too thick may reduce heat transfer efficiency and too thin may increase manufacturing cost.
In a more preferred embodiment, since the excitation light is irradiated from directly above the open container, the observation of the irradiated microdroplet is also directly above, and thus the field of view of the observation needs to be considered. The surrounding part 2 and the bottom plate part 1 form a certain included angle with each other, so that the opening area of the top of the open container is larger than the area of the bottom plate part 1, thus the observation area can be increased, and the influence of the observation field on the observation result can be reduced.
In a preferred embodiment, the use scenario of the digital PCR well plate is: the digital PCR well plate needs to be placed in a corresponding recess of the analysis platform of the fully automatic digital PCR analysis system. The top of the surrounding part 2 extends to the outside of the open container to form a rim part 9. When the digital PCR pore plate is placed in the full-automatic PCR analysis system, the edge part 9 can be used as a supporting platform, so that the digital PCR pore plate is convenient to use. The rim portion 9 may be formed by injection molding together when the open container is injection molded.
Further, as shown in fig. 3, since the digital PCR plate is designed symmetrically, in order to facilitate the user to quickly identify the front and back of the digital PCR plate, an identification notch 10 is provided at the edge 9, and the identification notch 10 is mainly used for determining the direction.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present utility model. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the utility model. Thus, the present utility model is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (11)
1. A digital PCR pore plate is characterized by comprising an injection-molded open container,
The open container has a bottom plate portion and a surrounding portion provided around an edge of the bottom plate portion; a plurality of flat bottom sample tanks which are sequentially arranged are arranged on the bottom plate part;
The roughness of the inner bottom surface of the flat bottom sample cell is 0.1-0.2 mu m.
2. The digital PCR well plate according to claim 1, wherein the roughness of the outer bottom surface of the open container is 0.02 μm to 0.06 μm.
3. The digital PCR well plate according to claim 1, wherein the bottom plate portion of the open container is provided with edge ribs and inner hole ribs; the edge reinforcing ribs are arranged around the bottom plate part, the inner hole reinforcing ribs are arranged inside the edge reinforcing ribs in a crisscross mode, and the inner space surrounded by the edge reinforcing ribs is divided into a plurality of flat bottom sample tanks.
4. The digital PCR well plate according to claim 3, wherein the flat bottom sample cell has a rectangular structure and the inner corners of the side walls of the flat bottom sample cell have circular arc structures.
5. The digital PCR well plate according to claim 3, wherein two strip-shaped waste liquid tanks are oppositely arranged on the edge reinforcing ribs in the open container.
6. The digital PCR well plate according to claim 3, wherein a plurality of linearly arranged glue reducing holes are further arranged on the inner edge reinforcing ribs of the open container; the gel reducing holes are positioned between two adjacent flat bottom sample tanks.
7. The digital PCR well plate of claim 1, further comprising a thermally conductive silicone sheet disposed at the outer bottom of the open container.
8. The digital PCR well plate according to claim 7, wherein the thermally conductive silicone sheet has a hardness of 30-50; the thickness of the heat conduction silica gel sheet is 0.5mm-1.5mm.
9. The digital PCR well plate according to claim 1, wherein the surrounding portion is obliquely disposed on the bottom plate portion.
10. The digital PCR well plate of claim 1, wherein the top of the surround extends outwardly to form a rim that is integrally formed with the open container.
11. The digital PCR well plate according to claim 10, wherein the rim portion is provided with an identification notch.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322049722.3U CN220887449U (en) | 2023-08-01 | 2023-08-01 | Digital PCR pore plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322049722.3U CN220887449U (en) | 2023-08-01 | 2023-08-01 | Digital PCR pore plate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN220887449U true CN220887449U (en) | 2024-05-03 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202322049722.3U Active CN220887449U (en) | 2023-08-01 | 2023-08-01 | Digital PCR pore plate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN220887449U (en) |
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2023
- 2023-08-01 CN CN202322049722.3U patent/CN220887449U/en active Active
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