Deep-hole plate suitable for high-throughput DNA extraction of large-volume seeds

Abstract

The utility model relates to the technical field of detection equipment, in particular to a deep hole plate suitable for high-flux DNA extraction of large-volume seeds, which is characterized in that a plurality of first specification holes and a plurality of second specification holes are formed in the deep hole plate; the first specification holes and the second specification holes are alternately arranged on the deep hole plate at intervals, the diameter of the first specification holes is larger than that of the second specification holes, and the first specification holes are used for containing large-volume seeds. And the compatibility with the existing automatic equipment can be kept.

Description

Technical Field

[0001] This utility model relates to the field of detection equipment technology, and in particular to a deep-well plate suitable for high-throughput DNA extraction from large-volume seeds. Background Technology

[0002] With the development of modern molecular breeding and gene research, the extraction and analysis of large-scale plant genomic DNA has become an important need in the field of biotechnology. Among many plant samples, DNA extraction from large-volume seeds such as maize has unique characteristics, which are not only reflected in the physical characteristics such as large seed volume and rigid tissue, but also in the special requirements for equipment and consumables when processing in batches.

[0003] Currently, standard consumables such as 96-well plates are commonly used in laboratories for DNA extraction. This design is primarily for small-volume samples such as leaves, and it has significant limitations when processing large-volume seeds. The pore size of a standard 96-well plate is typically less than 7 mm. This size is too small for large seeds such as corn, which have a diameter of 8-12 mm. This results in seeds not being able to be placed properly or, once placed, being unable to be processed due to limited space. Although the capacity problem can be solved by increasing the pore size, simply enlarging all pores will create new problems: First, the walls between adjacent wells become thinner, reducing the plate's strength and making it prone to cracking during agitation and grinding; second, using only the first-size pores will increase the amount of reagent needed for pipetting, resulting in reagent waste; third, using only the first-size pores will cause the overall size of the deep-well plate to exceed the standard specifications, making it incompatible with existing automated equipment.

[0004] Some laboratories have attempted to process large-volume seeds using larger centrifuge tubes or test tubes, but this approach is difficult to automate, and manual processing is not only inefficient but also prone to errors. Others have suggested using specially designed first-size deep-well plates, but their incompatibility with standard 96-channel pipetting workstations severely impacts experimental efficiency and ease of operation. Utility Model Content

[0005] This invention provides a deep-well plate suitable for high-throughput DNA extraction from large-volume seeds. This deep-well plate can meet the needs of large-volume seed processing while maintaining compatibility with existing automated equipment.

[0006] This utility model provides a deep well plate suitable for high-throughput DNA extraction from large-volume seeds. The deep well plate is provided with multiple first-size wells and multiple second-size wells, which are arranged alternately on the deep well plate. The diameter of the first-size wells is larger than that of the second-size wells, and the first-size wells are used to accommodate large-volume seeds.

[0007] In one possible implementation, the deep hole plate has 48 first-specification holes and 48 second-specification holes.

[0008] In one possible implementation, the center-to-center distance between adjacent first-specification holes and second-specification holes matches the nozzle spacing of a 96-channel high-throughput workstation.

[0009] In one possible implementation, the first specification hole and the second specification hole are arranged in 8 rows and 12 columns or 12 rows and 8 columns.

[0010] In one possible implementation, the deep hole plate includes a first deep hole plate and a second deep hole plate with identical structures, wherein a first specification hole on the first deep hole plate corresponds to a second specification hole on the second deep hole plate, and vice versa.

[0011] In one possible implementation, both the first-specification hole and the second-specification hole are cylindrical holes, with the diameter of the first-specification hole being 8-15 mm and the diameter of the second-specification hole being 3-6 mm.

[0012] In one possible implementation, the depth of the first specification hole is 15-30 mm.

[0013] In one possible implementation, the wall thickness between adjacent first-specification holes and second-specification holes is 2-4 mm.

[0014] In one possible implementation, the deep hole plate is made of polypropylene and has vertical reinforcing ribs on the inner walls of the first and second specification holes.

[0015] In one possible implementation, a sealing membrane is also included, which can detachably seal the first and second specification holes of the deep hole plate.

[0016] This invention provides a deep-well plate suitable for high-throughput DNA extraction from large-volume seeds. This deep-well plate effectively solves the problem of traditional 96-well plates being unable to accommodate large seeds such as corn by using a larger diameter first-size well to accommodate large seeds. The diameter of the first-size well is larger than that of the second-size well, providing ample space for large seeds. This design allows seeds to be smoothly placed into the wells while reserving sufficient space for soaking and swelling during DNA extraction. When large seeds such as corn are placed in the first-size well, appropriate gaps are left around them, allowing the extraction solution to fully wet the seed surface, improving DNA extraction efficiency. Furthermore, by employing an alternating arrangement design, smaller second-size wells are placed between the first-size wells, cleverly maintaining compatibility with 96-channel high-throughput workstations. Alternating the first and second-size wells on the deep-well plate ensures that each workstation's pipetting channel corresponds to a well position. The second-size well provides an independent, uncontaminated space for the pipetting tip to descend, while avoiding multiple pipetting channels corresponding to the same first-size well, ensuring the accuracy of automated operation. Operators can directly use the deep hole plate of this utility model for batch processing without making any modifications or adjustments to existing equipment. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0018] Figure 1 This is a top view schematic diagram of a first deep hole plate provided by this utility model.

[0019] Figure 2 This is a three-dimensional structural diagram of a first deep hole plate provided by this utility model.

[0020] Figure 3 yes Figure 2 A schematic diagram of the three-dimensional structure from another angle.

[0021] Figure 4 This is a top view of a second deep hole plate provided by this utility model.

[0022] Figure label:

[0023] 1. Deep hole plate; 11. First specification hole; 12. Second specification hole; 13. First deep hole plate; 14. Second deep hole plate; 15. Mistake-proof part. Detailed Implementation

[0024] To make the objectives, technical solutions, and advantages of this utility model clearer, the technical solutions of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model.

[0025] The following is combined Figure 1-4 This invention describes a deep-well plate 1 suitable for high-throughput DNA extraction from large-volume seeds. The deep-well plate 1 is provided with a plurality of first-size wells 11 and a plurality of second-size wells 12. The first-size wells 11 and the second-size wells 12 are arranged alternately on the deep-well plate 1. The diameter of the first-size wells 11 is larger than the diameter of the second-size wells 12. The first-size wells 11 are used to accommodate large-volume seeds.

[0026] In this invention, a larger-diameter first-size well 11 is used to accommodate large-volume seeds, effectively solving the problem that traditional 96-well plates cannot accommodate large-volume seeds such as corn. The diameter of the first-size well 11 is larger than that of the second-size well 12, providing ample space for large-volume seeds. This design allows seeds to be smoothly placed into the wells while reserving sufficient space for soaking and swelling during DNA extraction. When large-volume seeds such as corn are placed into the first-size well 11, appropriate gaps are left around them, allowing the extraction solution to fully wet the seed surface, improving DNA extraction efficiency. In addition, by adopting an alternating arrangement design, smaller-size second-size wells 12 are placed between the first-size wells 11, cleverly maintaining compatibility with 96-channel high-throughput workstations. The alternating arrangement of the first-size wells 11 and the second-size wells 12 on the deep-well plate 1 ensures that each workstation's pipetting channel corresponds to one well position. The second-size well 12 provides an independent space to prevent contamination when the pipetting tip descends, while avoiding the situation where multiple pipetting channels correspond to the same first-size well 11, ensuring the accuracy of automated operation. Operators can directly use the deep hole plate 1 of this utility model for batch processing without making any modifications or adjustments to existing equipment.

[0027] In related technologies, conventional DNA extraction methods combined with 96-channel high-throughput workstations, through programmed control of lysis, binding, and washing steps, reduce manual intervention and are currently the most commonly used DNA extraction method for large-scale sample testing, particularly for purity detection. However, different seeds and samples have different specifications, and the fixed pore size of traditional deep-well plates suitable for 96-channel high-throughput workstations is difficult to accommodate larger seed samples. Furthermore, corn seeds, being large and hard, present the following problems if soaked and ground using a 96-well plate:

[0028] 1. Insufficient seed capacity: Due to the limited size of each well in a 96-well plate, it is difficult to fully accommodate larger corn seeds, which may lead to the following three situations: First, the seeds are too large to be placed in the well; second, although the seeds can be placed in the well, they are stuck due to insufficient space, preventing the seeds from fully contacting the soaking reagent, affecting their swelling and softening effect, and thus reducing grinding efficiency; third, if the seeds are forcibly pressed into the well by external force, the compression between the seeds and the well wall may cause cracks in the well wall, increasing the risk of tube wall rupture during grinding, and thus causing cross-contamination of the samples.

[0029] 2. Insufficient space after seeds absorb water and swell: Corn seeds will absorb water and swell during soaking, and their volume will increase significantly. However, the space of the pores in the 96-well plate is limited and cannot accommodate the swollen seeds. This may result in the seeds not being able to fully absorb water and soften, thus affecting the subsequent grinding effect.

[0030] 3. Poor grinding effect: During the grinding process, insufficient space in the pores prevents the seeds from moving sufficiently, making it difficult for the steel balls to effectively strike the seeds, thus significantly reducing grinding efficiency. Furthermore, insufficient seed breakage further affects DNA release, reducing extraction efficiency.

[0031] In the embodiments provided by this utility model, the deep-hole plate 1, through its structural design of setting multiple first-size holes 11 and second-size holes 12 arranged alternately, effectively solves many problems encountered by single-size hole plates in the prior art when processing large-volume seeds. Specifically, the diameter of the first-size hole 11 is larger than that of the second-size hole 12, which can fully accommodate large-volume seeds such as corn, avoiding situations where seeds cannot be placed or are stuck due to their large size. In practical applications, when corn seeds are placed in the first-size hole 11, there is sufficient space for soaking and expansion, and the softening effect of the seeds is not affected by space limitations. At the same time, the larger space of the first-size hole 11 also provides sufficient room for steel ball grinding, improving grinding efficiency. The second-size hole 12, designed as a dummy hole, provides an independent space for the operation of the high-throughput workstation, avoiding the situation where multiple nozzles enter the same hole at the same time, reducing consumable waste. In addition, the alternating arrangement of large and small holes increases the hole spacing, improves the structural strength of the plate, and provides sufficient sealing area for the sealing film, effectively preventing cross-contamination of samples.

[0032] In some embodiments, the deep hole plate 1 is provided with 48 first specification holes 11 and 48 second specification holes 12.

[0033] In this invention, by setting a specific number of 48 first-specification wells 11 and 48 second-specification wells 12, the scheme achieves perfect compatibility with the standard 96-well plate. In practical applications, this design allows the deep-well plate 1 to fully utilize all channels of the 96-channel high-throughput workstation, ensuring that a single deep-well plate 1 can simultaneously process 48 large-volume seed samples while avoiding waste of equipment channels. This optimized well count design, while ensuring throughput, also provides sufficient structural support space for each first-specification well 11, ensuring the overall strength and stability of the plate. Furthermore, the 48+48 well count design facilitates the subsequent transfer of samples to the standard 96-well plate, making the entire experimental process more coherent and efficient.

[0034] In some embodiments, the center-to-center distance between adjacent first specification holes 11 and second specification holes 12 is matched with the nozzle spacing of a 96-channel high-throughput workstation.

[0035] In this invention, the center-to-center distance between adjacent first-specification holes 11 and second-specification holes 12 is matched with the spacing between the nozzles of the 96-channel high-throughput workstation, ensuring complete compatibility with existing automated equipment. In actual operation, this matching design allows the 96 nozzles of the workstation to precisely correspond to each hole, avoiding positioning deviations. Simultaneously, because each nozzle accurately enters its corresponding hole, operational errors are significantly reduced, improving the accuracy of sample processing. Furthermore, this design avoids the hassle of additional programming or parameter adjustments due to mismatched spacing, allowing the equipment to operate directly using standard procedures, thus improving experimental efficiency.

[0036] In some embodiments, the first specification hole 11 and the second specification hole 12 are arranged in 8 rows and 12 columns or 12 rows and 8 columns.

[0037] In this invention, the 8x12 or 12x8 arrangement ensures that the dimensions of the deep well plate 1 are consistent with those of the standard 96-well plate. This standardized design offers several advantages. First, the identical dimensions allow the deep well plate 1 to be placed directly on the work platform of existing experimental equipment without the need for additional adapters or supports. Second, this arrangement ensures optimal utilization of the plate space, providing sufficient space for each first-size well 11 while maintaining the overall structural compactness. Furthermore, the regular arrangement facilitates quick sample positioning by operators, reducing operational errors. In practical applications, this standardized design also facilitates the stacking and storage of multiple deep well plates 1, saving storage space.

[0038] In some embodiments, the deep hole plate 1 includes a first deep hole plate 13 and a second deep hole plate 14 with the same structure, wherein the first specification hole 11 on the first deep hole plate 13 corresponds to the second specification hole 12 on the second deep hole plate 14, and the second specification hole 12 on the first deep hole plate 13 corresponds to the first specification hole 11 on the second deep hole plate 14.

[0039] This invention cleverly solves the problem of processing large batches of samples by designing a first deep-well plate 13 and a second deep-well plate 14 with identical structures and making the positions of the first-specification holes 11 and the second-specification holes 12 of the two plates complementary. In practical applications, this design allows the two plates to process 96 large-volume seed samples simultaneously, significantly improving processing efficiency. Since the two plates have identical structures, they are interchangeable, simplifying laboratory inventory management. More importantly, the complementary hole positions of the two plates allow the processed samples to be completely transferred to a standard 96-well plate, ensuring the continuity of subsequent experimental procedures. This design also avoids the problem of needing multiple 96-well plates to accommodate large-volume samples, saving experimental consumables.

[0040] Specifically, the first deep hole plate 13 and the second deep hole plate 14 each have a foolproof part 15. The foolproof part 15 can be used to position the first deep hole plate 13 and the second deep hole plate 14, so that the first deep hole plate 13 and the second deep hole plate 14 can be used in a relative manner to introduce the sample in the first specification hole 11 into the standard 96 deep hole plate, which can ensure the smooth progress of subsequent high-throughput operations.

[0041] In some embodiments, the first specification hole 11 and the second specification hole 12 are both cylindrical holes, the diameter of the first specification hole 11 is 8-15mm, and the diameter of the second specification hole 12 is 3-6mm.

[0042] In this invention, the cylindrical orifice design and specific dimensional parameters provide an optimal solution for processing large-volume seeds. The first-specification orifice, with a diameter range of 118-15mm, ensures the capacity to accommodate corn seeds of various sizes while reserving sufficient space for seed expansion during soaking. The second-specification orifice, with a diameter of 123-6mm, perfectly meets the operational requirements of the pipette tip, ensuring smooth entry while avoiding reagent waste caused by excessive space. The cylindrical structure not only facilitates manufacturing but also ensures effective movement of the steel ball during grinding, improving grinding efficiency. Furthermore, this design also considers ease of cleaning and maintenance, as the cylindrical orifice eliminates cleaning dead zones.

[0043] In some embodiments, the depth of the first specification hole 11 is 15-30 mm.

[0044] In this invention, the 15-30mm hole depth design is determined based on practical application requirements. This depth range ensures sufficient space to accommodate the total volume of seeds, steel balls, and reagents, preventing liquid from splashing out during grinding. Simultaneously, this depth provides adequate space for the movement of the steel balls, allowing them to fully exert their grinding effect. In practical applications, this depth also considers the needs of pipetting operations, ensuring the pipette tip can smoothly reach the bottom of the hole for liquid aspiration, avoiding dead volume. Furthermore, this depth range balances the requirements of sheet strength and processing technology, ensuring structural strength without significantly increasing material costs.

[0045] Specifically, the depth of the second specification hole 12 can be the same as or less than the depth of the first specification hole 11, as long as it can accommodate the gun head of the workstation.

[0046] In some embodiments, the wall thickness between adjacent first specification holes 11 and second specification holes 12 is 2-4 mm.

[0047] In this invention, the 2-4mm partition wall thickness design represents the optimal balance between structural strength and space utilization of the deep-hole plate 1. This thickness range ensures sufficient strength between adjacent holes to withstand the stress during vibratory grinding, preventing hole wall cracking. Simultaneously, this thickness provides ample space for reinforcing ribs, further enhancing structural strength. In practical applications, this partition wall thickness also provides adequate thermal insulation, reducing temperature interference between adjacent holes. Furthermore, this thickness range also accommodates the sealing requirements of the membrane, providing sufficient sealing area and ensuring a good sealing effect.

[0048] In some embodiments, the deep hole plate 1 is made of polypropylene and has vertical reinforcing ribs on the inner walls of the first specification hole 11 and the second specification hole 12.

[0049] In this invention, the use of polypropylene material and the design of vertical reinforcing ribs on the inner wall of the borehole improves the performance of the deep-hole plate 1 in several ways. Polypropylene material has excellent chemical and temperature resistance, making it suitable for various extraction reagent environments and facilitating high-temperature sterilization. The vertical reinforcing ribs significantly improve the deformation resistance of the borehole wall, enabling it to withstand severe vibrations during grinding. In practical applications, this reinforced structure also prevents deformation of the borehole wall due to temperature changes, ensuring stable dimensions of the plate during long-term use. Furthermore, the reinforcing ribs improve the overall rigidity of the plate, reducing the risk of deformation during handling and operation.

[0050] In some embodiments, a sealing membrane is also included, which can detachably seal the first specification hole 11 and the second specification hole 12 of the deep hole plate 1.

[0051] In this invention, the design of a detachable sealing membrane and a sealing reinforcement structure effectively solves the sealing problem during sample processing. The detachable design allows the sealing membrane to be used multiple times as needed, saving on consumable costs. The sealing reinforcement structure between the sealing membrane and the upper surface of the deep-hole plate 1 provides a more reliable sealing effect, effectively preventing liquid leakage and cross-contamination during oscillation. In practical applications, this sealing design also facilitates the replacement of the sealing membrane between different experimental steps, improving operational flexibility. Furthermore, the sealing reinforcement structure increases the stress-bearing area of ​​the sealing membrane, extending its service life.

[0052] Specifically, by using first-specification holes 11 and second-specification holes 12 of different sizes, the hole spacing is larger, which enhances the sealing performance of the sealing film and prevents liquid from escaping and causing contamination between holes during vibration.

[0053] In summary, this utility model provides a deep-well plate 1 suitable for high-throughput DNA extraction from large-volume seeds. By setting a larger diameter first-size well 11 to accommodate large-volume seeds, it effectively solves the problem that traditional 96-well plates cannot accommodate large-volume seeds such as corn. The diameter of the first-size well 11 is larger than that of the second-size well 12, providing ample space for large-volume seeds. This design allows seeds to be smoothly placed into the wells while reserving sufficient space for soaking and swelling during DNA extraction. When large-volume seeds such as corn are placed into the first-size well 11, appropriate gaps are left around them, allowing the extraction solution to fully wet the seed surface, improving DNA extraction efficiency. In addition, by adopting an alternating arrangement design, and setting smaller second-size wells 12 between the first-size wells 11, compatibility with 96-channel high-throughput workstations is cleverly maintained. The alternating arrangement of the first-size wells 11 and the second-size wells 12 on the deep-well plate 1 ensures that each workstation's pipetting channel corresponds to one well. The second-specification hole 12 provides an independent, uncontaminated space for the pipetting tip to descend, while also preventing multiple pipetting channels from corresponding to the same first-specification hole 11, thus ensuring the accuracy of automated operation. Operators can directly use the deep-hole plate 1 of this invention for batch processing without any modification or adjustment to existing equipment.

[0054] The device embodiments described above are merely illustrative. The units described as separate components may or may not be physically separate. The components shown as units may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs. Those skilled in the art can understand and implement this without any creative effort.

[0055] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. A deep-well plate suitable for high-throughput DNA extraction from large-volume seeds, characterized in that, The deep hole plate (1) is provided with a plurality of first specification holes (11) and a plurality of second specification holes (12). The first specification holes (11) and the second specification holes (12) are arranged alternately on the deep hole plate (1). The diameter of the first specification hole (11) is larger than the diameter of the second specification hole (12). The first specification hole (11) is used to accommodate large-volume seeds.

2. The deep hole plate according to claim 1, characterized in that, The deep hole plate (1) is provided with (48) first specification holes (11) and (48) second specification holes (12).

3. The deep hole plate according to claim 2, characterized in that, The center-to-center distance between adjacent first specification holes (11) and second specification holes (12) matches the nozzle spacing of the (96) channel high-throughput workstation.

4. The deep hole plate according to claim 2, characterized in that, The first specification hole (11) and the second specification hole (12) are arranged in 8 rows and 12 columns or 12 rows and 8 columns.

5. The deep hole plate according to claim 1, characterized in that, The deep hole plate (1) includes a first deep hole plate (13) and a second deep hole plate (14) with the same structure, wherein the first specification hole (11) on the first deep hole plate (13) corresponds to the second specification hole (12) on the second deep hole plate (14), and the second specification hole (12) on the first deep hole plate (13) corresponds to the first specification hole (11) on the second deep hole plate (14).

6. The deep hole plate according to claim 1, characterized in that, Both the first specification hole (11) and the second specification hole (12) are cylindrical holes. The diameter of the first specification hole (11) is 8-15mm, and the diameter of the second specification hole (12) is 3-6mm.

7. The deep hole plate according to claim 6, characterized in that, The depth of the first specification hole (11) is 15-30mm.

8. The deep hole plate according to claim 1, characterized in that, The wall thickness between adjacent first specification hole (11) and second specification hole (12) is 2-4 mm.

9. The deep hole plate according to any one of claims 1-8, characterized in that, The deep hole plate (1) is made of polypropylene and has vertical reinforcing ribs on the inner walls of the first specification hole (11) and the second specification hole (12).

10. The deep hole plate according to any one of claims 1-8, characterized in that, It also includes a sealing membrane that can detachably seal the first specification hole (11) and the second specification hole (12) of the deep hole plate (1).

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