A CPG module for 1 fmol to 1 nmol synthesis yield

Abstract

A kind of 1fmol~1nmol synthesis yield CPG module, belong to DNA recombination technical field, including CPG carrier plate, the lower end surface of the CPG carrier plate is connected with distribution plate by bolt lock, the synthesis hole is opened in the CPG carrier plate, the CPG column is sintered in the synthesis hole, the distribution plate is opened in the liquid discharge channel, the synthesis hole and the liquid discharge channel are one to one;The CPG column is sintered by being screened into synthesis hole after CPG powder and ultra-high molecular weight polyethylene powder are premixed, and the lower end of the CPG column is flush with the lower end surface of the CPG carrier plate;First, modularization is convenient for later expansion and maintenance, without considering the economic loss caused by replacing the entire device due to the damage of a component, in addition, the CPG column is sintered in the synthesis hole, to avoid the problem that CPG column installation precision difference is large, leading to synthesis air resistance difference is large, improve synthesis efficiency and synthesis quality.

Description

Technical Field

[0001] This invention belongs to the field of DNA recombination technology, and specifically relates to a CPG module with a synthesis yield of 1 fmol to 1 nmol. Background Technology

[0002] DNA synthesis yield depends on the molar concentration of CPG, and the gas resistance of the CPG column is a crucial indicator that directly affects the synthesis quality. The gas resistance of CPG is influenced by the molar concentration and uniformity of the CPG packing. Most commercially available CPGs are manufactured by sintering the CPG carrier and then loading it into the synthesis module; this method is suitable for low-throughput applications.

[0003] However, there are serious problems with synthesis fluxes of 1K to 10K and above (referred to as high flux). Due to the small size of CPGs used in high flux synthesis, the CPG installation accuracy varies greatly, which affects the gas resistance of the CPGs, leading to an increase in gas resistance differences and ultimately affecting the synthesis efficiency and quality. Summary of the Invention

[0004] The purpose of this invention is to provide a CPG module with a synthesis yield of 1 fmol to 1 nmol to solve the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A CPG module with a synthesis yield of 1 fmol to 1 nmol includes a CPG carrier plate, the lower end of which is bolted to a separator plate. The CPG carrier plate includes synthesis holes, within which CPG columns are sintered. The separator plate has drainage channels, with each synthesis hole corresponding to one of the drainage channels. The CPG column is formed by premixing CPG powder and ultra-high molecular weight polyethylene powder, sieving the mixture into the synthesis holes, and then sintering the mixture. The lower end of the CPG column is flush with the lower end of the CPG carrier plate.

[0007] Compared with existing technologies, this technical solution has the following advantages:

[0008] First, modular design facilitates future expansion and maintenance without the need to consider the economic losses caused by replacing the entire unit due to the failure of a single component. In addition, by sintering the CPG column inside the synthesis borehole, the problem of large differences in CPG column installation accuracy leading to large differences in synthesis gas resistance is avoided, thereby improving synthesis efficiency and synthesis quality.

[0009] Preferably, the synthesis orifice includes an inlet section, a transition section, and a CPG carrier section arranged sequentially from top to bottom, which are connected in sequence, and the CPG column is sintered within the CPG carrier section.

[0010] Preferably, the CPG powder comprises a mixture of non-silanized CPG powder and silanized CPG powder.

[0011] Preferably, the inner diameter of the liquid inlet section is larger than that of the CPG carrier section, and the cross-section of the transition section in the vertical direction is an inverted trapezoid.

[0012] Preferably, the CPG carrier plate is made of high-density PP material.

[0013] Preferably, the inner surface of the synthetic pore is coated with a Teflon coating.

[0014] Preferably, the liquid separating plate includes a liquid separating plate body, the upper end face of the liquid separating plate body is provided with a liquid separating cavity, the lower end wall of the liquid separating cavity is provided with an array of drainage channels, and the lower end face of the liquid separating plate body is provided with a boss, the lower end of the drainage channel penetrates through the lower end face of the boss.

[0015] Preferably, a filter membrane is disposed between the liquid separator and the CPG carrier plate. In the assembled state, the filter membrane is sintered on the lower surface of the CPG carrier plate or the upper surface of the liquid separator; or, the lower surface of the filter membrane is pressed onto the upper surface of the liquid separator plate, and the upper surface of the filter membrane is pressed onto the lower surface of the CPG carrier plate.

[0016] Preferably, the CPG carrier plate includes a carrier plate body, and the upper end face of the carrier plate body is provided with threaded holes for accommodating bolts and waist-shaped grooves that are staggered with the threaded holes.

[0017] Preferably, the upper end face of the carrier plate body is recessed inward at the middle position, and the synthetic hole array is disposed in the recess. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the overall structure of the present invention;

[0019] Figure 2 yes Figure 1 A schematic diagram of the overall structure in the "A" direction;

[0020] Figure 3 This is an exploded view of the overall structure of the present invention;

[0021] Figure 4 yes Figure 3 A schematic diagram of the overall structure in the "B" direction;

[0022] Figure 5 This is a cross-sectional structural diagram of the present invention.

[0023] In the diagram: 1. CPG carrier plate; 10. Carrier plate body; 11. Synthesis hole; 110. Liquid inlet section; 111. Transition section; 112. CPG carrier section; a. CPG column; 12. Threaded hole; 13. Waist-shaped groove; 14. Cavity; 2. CPG separator plate; 20. Separator plate body; 200. Connecting hole; 21. Boss; 22. Drainage channel; 23. Separator chamber; 3. Bolt; 4. Filter membrane. Detailed Implementation

[0024] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0025] Example:

[0026] In this embodiment, CPG stands for controlled pore glass.

[0027] like Figure 3-4 The CPG module shown has a synthesis yield of 1 fmol to 1 nmol, and includes a CPG carrier plate 1, a filter membrane 4, and a separation plate 2 arranged from top to bottom.

[0028] In combination state, such as Figure 1-2 As shown, the CPG carrier plate 1 includes a carrier plate body 10. The upper end face of the carrier plate body 10 is recessed inward to form a cavity 14. The lower end face of the cavity 14 is provided with an array of synthesis holes 11. The edge of the upper end face of the carrier plate body 10 is provided with a plurality of threaded holes 12. Bolts 3 are placed within the threaded holes 12. The lower ends of the bolts 3 are connected to the distribution plate 2. That is, the distribution plate 2 has connection holes 200 at the positions where the bolts 3 are connected. (Refer to...) Figure 5 It can be seen that the threaded hole 12 and the connecting hole 200 are not as literal as they seem, that is, the carrier plate body 10 must be provided with a threaded hole 12 and the liquid distribution plate 2 must be provided with a connecting hole 200. They are only for better distinction and accurate expression.

[0029] Furthermore, the upper edge of the carrier plate body 10 is provided with a waist-shaped groove 13, which is staggered with the threaded hole 12. The advantages of providing the waist-shaped groove 13 are as follows: 1. Since the CPG carrier plate 1 in this embodiment is made by injection molding, it meets the technical requirements of uniform wall thickness of injection molding process; 2. The groove wall of the waist-shaped groove 13 constitutes the rib of the carrier plate, thereby strengthening the structural strength of the carrier plate itself.

[0030] It is worth noting that, in order to achieve better reaction of the reagents within the synthesis well 11 and subsequent cleaning, the synthesis well 11 comprises, from top to bottom, an inlet section 110, a transition section 111, and a CPG carrier section 112, which are connected sequentially. Specifically, the lower end of the inlet section 110 is connected to the upper end 111 of the transition section 111, and the lower end of the transition section 111 is connected to the upper end of the CPG carrier section 112. In this embodiment, the inlet section 110, the transition section 111, and the CPG carrier section 112 are integrally formed. Ideally, the inner surface formed by the three components should be an arc-shaped surface. However, due to the technical difficulties in real-world scenarios, the following solution was adopted: the inner diameter of the liquid inlet section 110 is larger than the inner diameter of the CPG carrier section 112, and the inner surface of the transition section 111 is a slope. The sloped structure of the transition section 111 facilitates injection molding and ensures dimensional stability. At the same time, connecting the liquid inlet section 110 and the CPG carrier section 112 achieves a buffering effect. Furthermore, the smooth surface avoids dead corners and facilitates reagent flow and cleaning.

[0031] Secondly, the CPG carrier segment 112 is provided with a CPG column a, for reference. Figure 3 and Figure 4 The CPG column a shown in the figure only indicates its placement within the synthesis orifice 11 and its corresponding positional relationship with the separatory plate 2. The specific relationship will be explained again below, therefore it will not be emphasized here. However, it should be noted that the CPG column a used in this embodiment is not pre-prepared, but rather prepared in advance through a sintering process. The specific preparation process is as follows:

[0032] (1) Preparation of raw materials: In this embodiment, 2000-pore CPG powder and ultra-high molecular weight polyethylene are selected as raw materials. The CPG powder is non-silanized CPG powder (abbreviated as blank CPG powder).

[0033] (2) Preparation: Silanized CPG powder is prepared by silanizing a portion of blank CPG powder. Then, according to the required molar concentration of CPG, the blank CPG powder and silanized CPG powder are premixed and then premixed with ultra-high molecular weight polyethylene to obtain mixed powder. The mixed powder is screened into CPG carrier section 112 by a vibrating screen and CPG column a is obtained by sintering process, thereby achieving the integrated structure requirement of CPG column a and CPG carrier plate 1.

[0034] Due to the small pore size, if the CPG column a is prepared in advance and filled in the CPG carrier segment 112, the operation requirements will be more stringent. Furthermore, external factors during installation will cause differences in the installation accuracy of the CPG column a in each synthesis well 11, resulting in differences in CPG gas resistance, which will ultimately affect the synthesis efficiency and quality of the DNA strand.

[0035] In this embodiment, the carrier plate body 10 is made of high-density PP material and is integrally manufactured by injection molding. This method ensures the stability of the manufacturing process and the economy of the manufacturing cost. In order to ensure the efficiency of subsequent cleaning and reduce the amount of cleaning reagent, a Teflon coating is sprayed on the inner surface of the synthesis hole 11 to reduce the phenomenon of synthesis reagent adhering to the wall.

[0036] Combination Figure 5 In this embodiment, the liquid distribution plate 2 includes a liquid distribution plate body 20. The upper end of the liquid distribution plate body 20 is recessed inward to form a liquid distribution cavity 23. The liquid distribution cavity 23 is provided with an array of drainage channels 22. In order to further improve the gas resistance performance of the product, a filter membrane 4 is provided in the liquid distribution cavity 23. The lower end of the synthesis hole 11 abuts against the upper end surface of the filter membrane 4.

[0037] Secondly, the lower end face of the liquid distribution plate 20 is provided with a boss 21. In actual application, the lower end face of the CPG module will be connected to a negative pressure system, which includes a negative pressure device (generally an air pump). The air extraction end pipe of the negative pressure device is connected to a waste liquid collection box. The liquid distribution plate 20 is inserted into the upper end face of the waste liquid collection box. In order to enhance the airtightness during insertion, a sealing ring is fitted on the outside of the boss 21. At the same time, in order to avoid the problem of splashing when the waste liquid flows from the liquid distribution plate 20 through the waste liquid collection box, the outlet end of the drain channel 22 extends into the waste liquid collection box. The edge of the liquid distribution plate 20 is provided with a connecting hole 200 for connection with the bolt 3, thereby realizing the locking of the CPG carrier plate 1 and the liquid distribution plate 2.

[0038] In the description of this invention, it should be understood that the terms "center," "lateral," "upper," "lower," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," and "outer," etc., indicating orientation or positional relationships based on the orientation or positional relationships shown in the accompanying drawings, are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, unless otherwise stated, "a number" means two or more. Additionally, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion.

[0039] This invention has been described with reference to embodiments. Without departing from the fundamental principles, several modifications and improvements can be made to this device. It should be noted that all technical solutions obtained through equivalent substitution or transformation fall within the protection scope of this invention.

Claims

1. A CPG module with a synthesis yield of 1 fmol to 1 nmol, comprising a CPG carrier plate (1), wherein a separatory plate (2) is bolted (3) to the lower end face of the CPG carrier plate (1); characterized in that: The CPG carrier plate (1) includes a synthesis hole (11) opened on the CPG carrier plate (1), and a CPG column (a) is sintered in the synthesis hole (11). A drain channel (22) is opened on the liquid distribution plate (2). The synthesis hole (11) and the drain channel (22) correspond one-to-one. The CPG column (a) is formed by premixing CPG powder and ultra-high molecular weight polyethylene powder, sieving it into the synthesis hole, and sintering it. The lower end of the CPG column (a) is flush with the lower end face of the CPG carrier plate (1). A filter membrane (4) is disposed between the liquid separator (2) and the CPG carrier plate (1). In the assembled state, the filter membrane (4) is sintered on the lower surface of the CPG carrier plate (1) or the upper surface of the liquid separator (2); or, the lower surface of the filter membrane (4) is pressed against the upper surface of the liquid separator (2), and the upper surface of the filter membrane (4) is pressed against the lower surface of the CPG carrier plate (1). The liquid distribution plate (2) includes a liquid distribution plate body (20), a liquid distribution cavity (23) is provided on the upper end face of the liquid distribution plate body (20), a drain channel (22) is arranged on the lower end wall of the liquid distribution cavity (23), and a boss (21) is provided on the lower end face of the liquid distribution plate body (20), and the lower end of the drain channel (22) penetrates the lower end face of the boss (21).

2. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 1, characterized in that: The synthesis orifice (11) includes a liquid inlet section (110), a transition section (111) and a CPG carrier section (112) arranged sequentially from top to bottom. The three sections are connected in sequence, and the CPG column (a) is sintered in the CPG carrier section (112).

3. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 2, characterized in that: The CPG powder comprises a mixture of non-silanized CPG powder and silanized CPG powder.

4. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 2, characterized in that: The inner diameter of the liquid inlet section (110) is larger than that of the CPG carrier section (112), and the cross-section of the transition section (111) in the vertical direction is an inverted trapezoid.

5. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 1, characterized in that: The CPG carrier plate (1) is made of high-density PP material.

6. A CPG module with a synthesis yield of 1 fmol to 1 nmol as described in any one of claims 1-5, characterized in that: The inner surface of the synthetic hole (11) is coated with a Teflon coating.

7. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 1, characterized in that: The CPG carrier plate (1) includes a carrier plate body (10), and the upper end face of the carrier plate body (10) is provided with a threaded hole (12) for accommodating a bolt (7) and a waist-shaped groove (13) intersecting with the threaded hole (12).

8. The CPG module with a synthesis yield of 1 fmol to 1 nmol as described in claim 7, characterized in that: The upper end face of the carrier plate body (10) is recessed inward at the middle position and a cavity (14) is provided. The array of synthetic holes (11) is arranged in the cavity (14).

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