CN116159608B

CN116159608B - A liquid dispensing device for a high-throughput gene synthesizer

Info

Publication number: CN116159608B
Application number: CN202211704039.2A
Authority: CN
Inventors: 张会刚; 郎秋蕾; 李璐璐; 肖晔
Original assignee: Hangzhou Lianchuan Biotechnology Co ltd
Current assignee: Hangzhou Lianchuan Biotechnology Co ltd
Priority date: 2022-12-29
Filing date: 2022-12-29
Publication date: 2025-11-04
Anticipated expiration: 2042-12-29
Also published as: CN116159608A

Abstract

A liquid addition device for a high-throughput gene synthesizer, belonging to the field of DNA recombination technology, includes a mounting plate with a mounting area. Liquid addition components are arrayed within the mounting area. Each liquid addition component includes a liquid addition base with liquid addition elements inserted into it. The liquid addition element has an outlet end for controlling the liquid flow rate. The liquid addition elements are arrayed on the liquid addition base. The array configuration allows for multiple combinations of liquid addition components to suit various application scenarios. Furthermore, the use of multiple liquid addition components significantly shortens the liquid addition time, thereby improving synthesis efficiency.

Description

Liquid feeding device of high flux gene synthesizer

Technical Field

The invention belongs to the technical field of DNA recombination, and particularly relates to a liquid adding device of a high-flux gene synthesizer.

Background

At present, the artificially synthesized DNA is the only way for the known directional modification of gene sequences, and is widely applied to a plurality of fields such as protein modification, life science and the like, such as nucleic acid medicine, enzyme engineering, gene detection, gene therapy and the like.

Most DNA synthesizers in the current market adopt phosphoramidite chemical synthesis to synthesize DNA, and the DNA synthesis is carried out according to four steps of deprotection, activation coupling, capping and oxidation, wherein different chemical reagents are needed in each step. Wherein the deprotection reagent is mainly trichloroacetic acid solution (TCA), the four monomer reagents are T, G and C.A, the activating reagent is ACT, the capping reagent is CAPA and CAPB, the oxidizing reagent is mainly iodine solution (Ox), and the cleaning reagent is mainly acetonitrile solution (ACN). Conventional porous synthetic plates are typically either 8 x12 wells (96-throughput) or 16 x 24 wells (384-throughput), and during the synthesis process, after the synthetic support (typically a CPG column) is pre-loaded into each well on each synthetic plate, the synthetic plate is placed into the instrument to begin synthesis, and the corresponding reagent is added to each well to ensure that the reagent reacts well with the support in each synthetic well in the synthetic plate. For a 96-flux or 384-flux synthesizer, a plurality of liquid adding spray heads are used for simultaneously spraying liquid into a row of holes, most of synthesizers are divided into 8 or 16 liquid adding spray heads for adding liquid for each reagent, the distance between the spray heads is 9mm, and the liquid spraying times are 12 or 48.

However, compared with a 1536-flux synthesizer, the synthetic plate has 32×48 holes, the hole-to-hole spacing is 2.25mm, if each reagent still uses a 9 mm-spaced nozzle to add liquid, the liquid adding module needs to run back and forth four times to add 1536 holes with the reagent, which results in the problems of too long liquid adding time, low synthesis efficiency, and the like. Therefore, for high throughput synthesizers, a high density of the liquid injection nozzles is highly desirable to address this problem.

Disclosure of Invention

The invention aims to provide a liquid adding device of a high-flux gene synthesizer, which aims to solve the problems in the background technology.

In order to achieve the above purpose, the present invention provides the following technical solutions:

the liquid adding device of the high-flux gene synthesizer comprises a mounting plate, wherein a mounting area is arranged on the mounting plate, a liquid adding assembly is arranged in the mounting area in an array mode, the liquid adding assembly comprises a liquid adding base, a liquid adding piece is inserted into the liquid adding base, a liquid outlet piece for controlling liquid outlet amount is arranged at a liquid outlet end of the liquid adding piece, and the liquid adding piece array is arranged on the liquid adding base.

Compared with the prior art, the technical scheme has the following effects:

The mode that adopts the array realizes the many specifications combination of liquid feeding subassembly to be applicable to multiple scene demand, simultaneously, owing to set up the prerequisite of a plurality of liquid feeding subassemblies, show the time of shortening the liquid feeding, thereby promote synthetic efficiency.

Preferably, the liquid adding member is a liquid adding tube.

Preferably, the liquid outlet piece is a steel needle.

Preferably, the liquid outlet end of the steel needle is subjected to reaming treatment, and the outer wall surface of the steel needle is coated with a Teflon coating.

Preferably, the liquid adding base is detachably connected with a clamping block through a bolt, a premixing cavity is arranged in the clamping block, two liquid adding pipes are arranged on the clamping block in an interference mode, gaps exist between the liquid outlet ends of the liquid adding pipes and the bottom end faces of the premixing cavity, the liquid inlet ends of the liquid outlet pieces are inserted on the clamping block, and the liquid inlet ends of the liquid outlet pieces are flush with the bottom end faces of the premixing cavity.

Preferably, the liquid adding base is provided with a plurality of liquid adding pipes in an array manner, and the liquid outlet ends of the liquid adding pipes are respectively provided with a steel needle in an inserting manner.

Drawings

FIG. 1 is a schematic view of the overall structure of the present invention;

FIG. 2 is a schematic view of the overall structure of the first liquid adding component in the invention;

FIG. 3 is an enlarged schematic view of a first liquid feeding assembly of the present invention, partially in section;

FIG. 4 is a first schematic view of the overall structure of the second liquid feeding assembly of the present invention;

FIG. 5 is a second schematic view of the overall structure of the second liquid feeding assembly of the present invention;

FIG. 6 is a schematic cross-sectional view of a second liquid feeding assembly according to the present invention.

Detailed Description

The following description of the embodiments of the present invention will be made with reference to the accompanying drawings, in which it is evident that the embodiments described are only some embodiments of the present invention, but not all embodiments.

Examples:

The liquid adding device of the high-flux gene synthesizer as shown in fig. 1-6 comprises a mounting plate 1, wherein a mounting area 10 is arranged on the mounting plate 1, a liquid adding component is arranged in an array in the mounting area 10, the liquid adding time is obviously shortened by increasing the density of the liquid adding component, the synthesis efficiency is improved, the liquid adding component comprises a liquid adding base, a liquid adding piece is arranged on the liquid adding base in an array, a liquid outlet piece for controlling the liquid outlet amount is arranged at the liquid outlet end of the liquid adding piece, and more specifically, in the embodiment, the liquid adding piece and the liquid outlet piece are respectively a liquid adding pipe and a steel needle, compared with the existing liquid adding device, in the embodiment, the liquid adding component is provided with two types;

For a better understanding of the structure of the liquid feeding assembly in this embodiment, the following will explain the structures of two types of liquid feeding assemblies in this embodiment in detail:

The first type is a single tube liquid adding mode, in this embodiment, the first liquid adding mechanism 3 is abbreviated as a first liquid adding mechanism 3, the first liquid adding mechanism 3 includes a first liquid adding support 30, a rod body of the first liquid adding support 30 is provided with an installation area 31, the installation area 31 is provided with an installation groove 310 in an array, a first liquid adding tube 33 is inserted into the installation groove 310, in order to enhance stability of the first liquid adding tube 33 during installation, a reinforcing block 35 is sleeved on a tube body of the first liquid adding tube 33 accommodated in the installation groove 310, meanwhile, for overall aesthetic property, an upper end face of the installation area 31 is connected with a sealing cover 32 through a bolt, an opening of the installation groove 310 is concealed, and secondly, a tube body of the first liquid adding tube 33 passes through a cover body of the sealing cover 32, as can be seen in combination with fig. 6, a liquid outlet end of the first liquid adding tube 33 is inserted with a first steel needle 34, and two ends of the first steel needle 34 are hollow for guiding out a reagent in the first liquid adding tube 33.

The second type is a double-tube liquid adding mode, in this embodiment, the second liquid adding mechanism 2 includes a second liquid adding support 20, an installation area 21 is provided on the second liquid adding support 20, the installation area 21 is provided with a plurality of clamping blocks 22 in an array, the clamping blocks 22 are detachably installed in the installation area 21 through bolts 220, two inserting holes are provided on the clamping blocks 22, a second liquid adding tube 23 is inserted into the inserting holes, it should be noted that a premix chamber is hollow in the installation area 21, that is, the installation area 21 corresponding to the downward extension of the clamping blocks 22 is provided with the premix chamber, and the purpose of the arrangement is that if multiple reagents are needed, compared with the single-tube liquid adding mode, multiple liquid injection needs to be performed, the mode can achieve simultaneous liquid injection, and meanwhile, as the multiple reagents can be subjected to certain premixing in the premix chamber, the coupling between the reagents is improved, and the premix chamber is internally provided with a sealing ring, so that the mixed reagents in the premix chamber are prevented from leaking, and the clamping blocks 22 are inserted into the second needle 24 to the second needle 24, and the second needle 24 extends to the second needle 24.

In this embodiment, the first type or the second type, or even the combination of the first type and the second type, is adopted, the purpose of high-flux rapid liquid adding is achieved and solved by adopting an array mode, meanwhile, the first steel needle 34 and the second steel needle 24 are both made of medical stainless steel, liquid outlet ends of the first steel needle and the second steel needle are both subjected to reaming treatment, and teflon coatings are sprayed on the outer walls to solve the problem of hanging drops after liquid adding;

and secondly, the combination can be realized according to actual requirements by adopting a detachable mode of bolt connection, so that the flexibility of the device is improved.

In the description of the present invention, it should be understood that the terms "center," "lateral," "upper," "lower," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like indicate or are based on the orientation or positional relationship shown in the drawings, merely to facilitate description of the invention and simplify the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, unless otherwise indicated, the meaning of "a number" is two or more. In addition, the term "include" and any variations thereof are intended to cover a non-exclusive inclusion.

The invention has been described in terms of embodiments, and the device can be modified and improved without departing from the principles of the invention. It should be noted that all technical solutions obtained by equivalent substitution or equivalent transformation fall within the protection scope of the present invention.

Claims

1. A liquid dispensing device for a high-throughput gene synthesizer, comprising a mounting plate (1), wherein the mounting plate (1) is provided with a mounting area (10), characterized in that: a liquid dispensing assembly is arrayed within the mounting area (10); the liquid dispensing assembly comprises a liquid dispensing base, wherein a liquid dispensing element is inserted into the liquid dispensing base, wherein the liquid dispensing element has a liquid outlet end provided with a liquid outlet element for controlling the liquid dispensing volume, and the liquid dispensing element is arrayed on the liquid dispensing base;

A clamping block (22) is detachably connected to the liquid adding base by bolts (220). A premixing chamber is hollow inside the liquid adding base corresponding to the clamping block (22). Two liquid adding pipes are interference-fitted on the clamping block (22). There is a gap between the liquid outlet end of the two liquid adding pipes and the bottom end face of the premixing chamber. The liquid inlet end of the liquid outlet is inserted into the clamping block (22). The liquid inlet end of the liquid outlet is flush with the bottom end face of the premixing chamber.

2. The liquid addition device for a high-throughput gene synthesizer as described in claim 1, wherein the liquid addition component is a liquid addition tube.

3. The liquid dispensing device for a high-throughput gene synthesizer as described in claim 2, wherein the liquid dispensing component is a steel needle.

4. The liquid dispensing device for a high-throughput gene synthesizer as described in claim 3, characterized in that: the liquid outlet end of the steel needle is enlarged, and the outer wall surface of the steel needle is coated with a layer of Teflon coating.

5. The liquid addition device for a high-throughput gene synthesizer as described in claim 3 or 4, characterized in that: a plurality of liquid addition tubes are arrayed on the liquid addition base, and steel needles are inserted into the liquid outlet ends of the plurality of liquid addition tubes.