Sheet gene synthesis substrate with high throughput and low reagent dosage
By designing a thickened skirt body structure on the thin-sheet DNA synthesis substrate, the problem of light and thinness of the substrate structure is solved, and gene synthesis with high throughput and low reagent dosage is achieved, ensuring synthesis accuracy and structural stability.
Patent Information
- Application Number
- CN202421660407.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-07-15
- Publication Date
- 2025-05-09
- Estimated Expiration
- 2034-07-15
AI Technical Summary
The existing thin-sheet DNA synthesis substrate structure is too thin, difficult to form stable installation, and is prone to deformation, which cannot meet the needs of high-throughput gene synthesis.
The thickened skirt body structure is fixed to reduce the stress on the base plate and prevent deformation of the base plate. The specific implementation is that the base plate includes a body and a skirt body, with a plurality of micro-holes arranged through the body, and the skirt body extends in the thickness direction from the outer circumference of the body, extending beyond the first and/or the second surface of the body, and the solid phase synthesis carrier is fixed in the micro-hole one by one.
Effectively prevent substrate plate deformation, improve structural strength and flatness, ensure synthesis accuracy, and achieve high throughput and low reagent dosage gene synthesis.
Smart Images
Figure CN222846710U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of gene synthesis, in particular to a thin-sheet gene synthesis substrate with high throughput and low reagent dosage. Background Art
[0002] DNA synthesis is one of the core tools in the upstream of synthetic biology and is crucial to promoting the development of downstream platforms and applications of synthetic biology. Among the current DNA synthesis technologies, column synthesis technology, as the first generation of synthesis technology, has the advantages of being the earliest developed and mature in technology, and has been widely used in the field of primer synthesis. However, with the development of synthetic biology, especially the increasing demand for high-throughput gene synthesis, traditional column synthesis technology can no longer fully meet this demand.
[0003] At present, in order to meet the needs of high-throughput gene synthesis, a thin-sheet DNA synthesis substrate is used, which includes a microwell plate and a solid-phase synthesis carrier. The microwell plate is provided with a large number of microwells for synthesis, and a solid-phase synthesis carrier is fixed in each microwell. However, the structure of this thin-sheet DNA synthesis substrate is too thin and light, and it is not easy to form a stable installation, and it is easy to deform. Utility Model Content
[0004] Based on this, it is necessary to provide a high-throughput, low-reagent-use, thin-sheet gene synthesis substrate that can be fixed by a thickened skirt structure to reduce the stress on the substrate and effectively prevent deformation of the substrate, in order to address the problem that the synthetic substrate structure is thin and easy to deform.
[0005] A high-throughput, low-reagent-usage thin-sheet gene synthesis substrate, comprising:
[0006] A base plate, comprising a main body and a skirt body, wherein the main body is provided with a plurality of micro-holes penetrating therethrough, the main body comprises a first surface and a second surface along a thickness direction, the skirt body is formed by extending the outer periphery of the main body along the thickness direction, and the skirt body extends beyond the first surface and / or beyond the second surface; and
[0007] The number of solid phase synthesis carriers corresponds to the number of the micropores, and the solid phase synthesis carriers are fixed in the micropores in a one-to-one correspondence.
[0008] In the above-mentioned high-throughput and low-reagent-usage thin-sheet gene synthesis substrate, each solid-phase synthesis carrier is fixed in each micropore of the body for gene synthesis. When positioning and installing the substrate plate, it can be fixed and supported by the thickened skirt body on the periphery of the body, which effectively reduces the force on the body. The skirt body is thicker than the body in the synthesis area, which not only strengthens the overall structural strength of the substrate plate, but also ensures the overall flatness of the body, effectively preventing the body from deforming and affecting the synthesis accuracy.
[0009] In one embodiment, the skirt body exceeds the first surface and the second surface respectively.
[0010] In one embodiment, the thickness of the skirt body beyond the first surface is less than the thickness beyond the second surface.
[0011] In one of the embodiments, the skirt body extends along the circumferential outer edge of the body.
[0012] In one embodiment, the skirt body is in the shape of a ring connected end to end.
[0013] In one embodiment, the microhole is a straight cylindrical through hole or a polygonal through hole.
[0014] In one embodiment, the port area of the micropore close to the first surface is larger than the port area close to the second surface.
[0015] In one embodiment, the micropore is conical; or the micropore is a stepped hole, a step is formed in the middle of the micropore, and the cross-sectional width of one end of the micropore close to the first surface is greater than the cross-sectional width of the other end close to the second surface.
[0016] In one embodiment, the plurality of micro-holes are arranged in an array or in a staggered arrangement.
[0017] In one embodiment, the plurality of micro-holes are arranged in an array, and the body is provided with a plurality of recessed grooves in the transverse direction and / or the longitudinal direction, and the depth of the grooves is less than the thickness of the body. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 This is a schematic structural diagram of an embodiment of a thin sheet gene synthesis substrate of the utility model;
[0019] Figure 2 This is a schematic cross-sectional structure diagram of a thin sheet gene synthesis substrate of the utility model;
[0020] Figure 3 for Figure 2 A local enlarged view of point A shown in FIG.
[0021] Figure 4 This is a structural schematic diagram of the first embodiment of the base plate of the utility model;
[0022] Figure 5 This is a structural schematic diagram of the second embodiment of the base plate of the utility model;
[0023] Figure 6 This is a structural schematic diagram of the third embodiment of the base plate of the utility model;
[0024] Figure 7This is a schematic structural diagram of a first embodiment of the micropore of the utility model;
[0025] Figure 8 This is a schematic structural diagram of the second embodiment of the micropore of the utility model;
[0026] Fig. 9 This is a structural schematic diagram of a fourth embodiment of the base plate of the utility model;
[0027] Fig.10 It is a schematic cross-sectional structure diagram of a base plate of the utility model.
[0028] In the accompanying drawings, the components represented by the reference numerals are listed as follows:
[0029] 100. Thin sheet gene synthesis substrate; 1. Matrix plate; 11. Main body; 111. First surface; 112. Second surface; 113. Groove; 12. Skirt body; 13. Micropore; 2. Solid phase synthesis carrier. DETAILED DESCRIPTION
[0030] In order to make the above-mentioned purposes, features and advantages of the utility model more obvious and easy to understand, the specific implementation methods of the utility model are described clearly and completely below in conjunction with the accompanying drawings. Obviously, the specific details described below are only part of the embodiments of the utility model, and the utility model can also be implemented in many other embodiments different from those described here. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the protection scope of the utility model.
[0031] It should be noted that when an element is referred to as being "fixed to" another element, it may be directly on the other element or there may be a central element. When an element is considered to be "connected to" another element, it may be directly connected to the other element or there may be a central element at the same time. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only and do not represent the only implementation method.
[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.
[0033] See also Figures 1 to 3In one embodiment, a high-throughput, low-reagent thin-sheet gene synthesis substrate 100 includes: a substrate plate 1 and a solid-phase synthesis carrier 2. The substrate plate 1 includes a body 11 and a skirt body 12. The body 11 is provided with a plurality of micropores 13 that penetrate therethrough. The body 11 includes a first surface 111 and a second surface 112 along the thickness direction. The skirt body 12 is formed by extending the periphery of the body 11 along the thickness direction. The skirt body 12 extends beyond the first surface 111 and / or beyond the second surface 112. The number of solid-phase synthesis carriers 2 corresponds to the number of micropores 13, and the solid-phase synthesis carriers 2 are fixed in the micropores 13 one by one. The thickness of the body 11 is less than 1 mm, the diameter of the solid-phase synthesis carrier 2 is less than 1.5 mm, and the reagent dosage is less than 1 μl.
[0034] In the high-throughput, low-reagent-usage thin-sheet gene synthesis substrate 100, each solid-phase synthesis carrier 2 is fixed in each microwell 13 of the body 11 for gene synthesis. When positioning and installing the base plate 1, the thickened skirt body 12 on the outer periphery of the body 11 can be used for fixing and supporting, which effectively reduces the force on the body 11. The skirt body 12 is thicker than the body 11 in the synthesis area, which not only strengthens the overall structural strength of the base plate 1, but also ensures the overall flatness of the body 11, effectively preventing the body 11 from deforming and affecting the synthesis accuracy.
[0035] In one embodiment in which the solid-phase synthesis carrier 2 is fixed in the micropores 13, the base plate 1 is first placed in the inner cavity of the sintering device and fixed, and the raw material powder to be sintered into the carrier is evenly spread into each micropore 13 of the body 11. Then, the raw material powder of the synthetic carrier is sintered into the solid-phase synthesis carrier 2 by sintering, and the solid-phase synthesis carrier 2 and the micropores 13 of the body 11 are adhered to form a whole, which is collectively called a gene synthesis substrate.
[0036] In the structural embodiment of the body 11, the substrate 1 as a whole can be a flat substrate made of corrosion-resistant glass, ceramics, silicon wafers, polypropylene PP, polyethylene PE, fluorinated ethylene propylene copolymer FEP and polytetrafluoroethylene PTFE, etc., and processed into micropores 13 of required diameters through machining, laser, etching, 3D printing and other processes. Figure 4 and Figure 5 The micropores 13 can be arranged in an array or in a staggered arrangement. The number of micropores 13 of the substrate plate 1 is determined by the area of the printed substrate, and a plurality of micropores 13 can be used as a standard. The staggered arrangement can increase the synthetic flux by more than 1.15 times under the same spacing. The surface of the substrate plate 1 is smooth, without barbs, frosting, or bends, with uniform flatness, and the appearance shape can be a rectangular flat plate, a circular flat plate, or other geometric shapes.
[0037] The main components of the raw materials of the solid phase synthesis carrier 2 include CPG powder and a bonding polymer material, wherein a DMT-protected or unprotected OH-containing connector is connected to the CPG, which is used as the starting point of the synthesis to connect the first phosphoramidite base. The CPG powder and the bonding polymer material are mixed in a certain proportion and sintered at a high temperature to form a solid phase synthesis carrier 2.
[0038] In the embodiment of the micropore 13 structure of the body 11, the micropore 13 can be a straight through hole or a polygonal through hole. The processing method of this direct through-type micropore 13 is simple and fast, with low processing difficulty and low cost.
[0039] See also Figure 6 In addition, in order to fix the solid phase synthesis carrier 2 in one direction and prevent it from falling and stably installing it so that it is not easy to shake, the micropores 13 can also be arranged so that the port area close to the first surface 111 is larger than the port area close to the second surface 112. Figure 7 Further, the micropore 13 may be tapered, or the micropore 13 may be a stepped hole, see Figure 8 A step is formed in the middle of the micropore 13, and the cross-sectional width of the end of the micropore 13 close to the first surface 111 is greater than the cross-sectional width of the end close to the second surface 112. The step hole can ensure that the solid phase synthesis carrier 2 is fixed while increasing the contact area and improving the adhesion.
[0040] In the embodiment of the skirt body 12, the skirt body 12 extends in the thickness direction of the body 11 and exceeds the first surface 111 and the second surface 112 respectively, forming a stable support for the body 11. The thickness of the skirt body 12 exceeding the first surface 111 is less than the thickness exceeding the second surface 112, which facilitates the synthesis operation from the first surface 111 to the micropore 13 while ensuring structural stability.
[0041] Furthermore, the skirt body 12 extends along the outer circumferential edge of the body 11, and can better provide protection for the body 11 and structural support on the plane when the base plate 1 is installed. The skirt body 12 can be annular and connected end to end, providing structural support and protection for the body 11 to the greatest extent.
[0042] See also Fig. 9 and Fig.10 After the synthesis of the substrate 1 is completed, the synthetic products can be sorted by cutting with a knife. Of course, it is also possible that when multiple micropores 13 are arranged in an array, multiple recessed grooves 113 are provided on the body 11 in the horizontal and / or vertical direction, and the depth of the grooves 113 is less than the thickness of the body 11. After the synthesis of the substrate 1 with pre-processed grooves 113, the substrate 1 can be quickly and easily sorted into several groups with the help of external force for subsequent product processing.
[0043] Processing technology of thin-sheet gene synthesis substrate 100: Step 1, place the substrate plate 1 in the inner cavity of the sintering device and lay it flat and fix it, without bending, bending, warping, etc.; Step 2, use a special scraper to evenly fill the solid-phase synthesis carrier 2 raw material into the micropores 13 of the substrate plate 1, lay it evenly in the multiple micropores 13, and compact and scrape it flat to fix it, to ensure that there is no missing or damage after sintering in the multiple micropores 13, and there is no solid-phase synthesis carrier 2 raw material adhering to the outside of the micropores 13, to prevent the reagent from flowing into other micropores 13, consuming too much reagent, and affecting the synthesis yield of adjacent micropores 13. Step 3, seal the sintering device, seal it to prevent air leakage, and turn the raw material into a solid-phase synthesis carrier 2 finished product through sintering, and form adhesion with the substrate plate 1, and sintering is completed. The thin-sheet gene synthesis substrate 100 can be directly used for gene synthesis.
[0044] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0045] The above-mentioned embodiments only express several implementation methods of the utility model, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the utility model patent. It should be pointed out that for ordinary technicians in this field, several deformations, substitutions and improvements can be made without departing from the concept of the utility model, which should be included in the protection scope of the utility model. Therefore, the protection scope of the utility model patent should be based on the claims.
Claims
1. A high-throughput, low-reagent-usage thin-sheet gene synthesis substrate, characterized in that: include: A base plate, comprising a main body and a skirt body, wherein the main body is provided with a plurality of micro-holes arranged therethrough, the main body comprises a first surface and a second surface along a thickness direction, the skirt body is formed by extending the outer periphery of the main body along the thickness direction, and the skirt body extends beyond the first surface and / or beyond the second surface; and The number of solid phase synthesis carriers corresponds to the number of the micropores, and the solid phase synthesis carriers are fixed in the micropores in a one-to-one correspondence.
2. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 1, characterized in that: The skirt body exceeds the first surface and the second surface respectively.
3. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 2, characterized in that: The thickness of the skirt body beyond the first surface is smaller than the thickness beyond the second surface.
4. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 2, characterized in that: The skirt body extends along the circumferential outer edge of the main body.
5. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 4, characterized in that: The skirt body is in the shape of a ring connected end to end.
6. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 1, characterized in that: The micropores are straight cylindrical through holes or polygonal through holes.
7. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 1, characterized in that: The port area of the micropore close to the first surface is larger than the port area close to the second surface.
8. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 7, characterized in that: The micropore is conical; or the micropore is a stepped hole, a step is formed in the middle of the micropore, and the cross-sectional width of one end of the micropore close to the first surface is greater than the cross-sectional width of one end close to the second surface.
9. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 1, characterized in that: The plurality of micropores are arranged in an array or in a staggered arrangement.
10. The high-throughput, low-reagent-usage thin-sheet gene synthesis substrate according to claim 1, characterized in that: The plurality of micro-holes are arranged in an array, and the body is provided with a plurality of recessed grooves in a transverse direction and / or a longitudinal direction, and the depth of the grooves is less than the thickness of the body.