CN113559803A

CN113559803A - DNA synthesis device and synthesis method

Info

Publication number: CN113559803A
Application number: CN202110937388.8A
Authority: CN
Inventors: 马石金; 朱强; 杜军; 陈园园; 周志康
Original assignee: Beijing Tsingke Biotech Co Ltd
Current assignee: Beijing Tsingke Biotech Co Ltd
Priority date: 2021-08-16
Filing date: 2021-08-16
Publication date: 2021-10-29
Anticipated expiration: 2041-08-16
Also published as: CN113559803B

Abstract

The invention relates to a DNA synthesis device and a synthesis method. The DNA synthesis device comprises: a synthesis column, a reaction cavity is formed inside the synthesis column, and the reaction cavity is used for accommodating a nucleic acid synthesis carrier; a reagent input component, the The reagent input assembly is used for inputting reagents into the reaction chamber; the intake assembly includes an intake member and a gas source, and the gas source is used for supplying gas, and the intake member is connected to the synthesis column. connection, the gas enters the reaction chamber through the air inlet to oscillate the nucleic acid synthesis carrier and the reagent in the reaction chamber; the reagent discharge component is used for removing the reaction chamber The reagents that end the reaction in the body are expelled. The DNA synthesis device can make the nucleic acid synthesis carrier more fully react with the reagent, thereby improving the synthesis rate.

Description

DNA synthesis device and synthesis method

Technical Field

The present invention relates to the technical field of DNA synthesis, and particularly to a DNA synthesis apparatus and a DNA synthesis method.

Background

The artificial synthesis of DNA is the only way of directionally modifying gene sequences known at present, and is widely applied to various fields of protein modification and life science, such as nucleic acid medicine, enzyme engineering, gene therapy and the like. When synthesizing, the common method is to put the nucleic acid synthesis carrier into the synthesis column, and sequentially introduce different reagents into the synthesis column, so that the nucleic acid synthesis carrier sequentially reacts with various reagents to obtain the final product. However, in the related art, when some DNA synthesis apparatuses perform DNA synthesis, the nucleic acid synthesis carrier cannot sufficiently react with the reagent, and the synthesis rate is low.

Disclosure of Invention

Based on this, the present invention provides a DNA synthesizer capable of improving the synthesis rate by allowing a nucleic acid synthesis vector to react with a reagent more sufficiently.

A DNA synthesis apparatus comprising:

the device comprises a synthesis column, a reaction cavity and a reaction cavity, wherein the reaction cavity is formed inside the synthesis column and is used for accommodating a nucleic acid synthesis carrier;

a reagent input assembly for inputting a reagent into the reaction chamber;

the gas inlet component comprises a gas inlet part and a gas source, the gas source is used for providing gas, the gas inlet part is connected with the synthesis column, and the gas enters the reaction cavity through the gas inlet part so as to oscillate the nucleic acid synthesis carrier and the reagent in the reaction cavity;

and the reagent discharging assembly is used for discharging the reagent which finishes the reaction in the reaction cavity.

In one embodiment, the connection position of the air inlet and the synthesis column is positioned at one side of the reaction cavity, and the reagent input assembly is positioned above the reaction cavity.

In one embodiment, the synthetic column comprises a main body part and a protruding part, the protruding part protrudes outwards from one side of the main body part, the reaction cavity is formed inside the main body part, the protruding part is hollow and communicated with the reaction cavity, and the air inlet part is connected with the protruding part.

In one embodiment, the air inlet extends into the reaction cavity from the protruding part to the radial center position.

In one embodiment, the inner diameter of the gas outlet channel of the inlet is less than 2mm, and may be, for example, 1.55mm, 0.84mm, 0.61mm, 0.26mm, 0.10mm or 0.06 mm.

In one embodiment, the air inlet piece comprises a luer connector and an air inlet needle, the luer connector is fixedly connected with the air inlet needle, and the air inlet needle is inserted into the protruding portion and is in interference fit with the protruding portion.

In one embodiment, the reagent discharging assembly is connected to the synthesis column, the reagent discharging assembly comprises a suction piece, and a filter element is arranged in the reaction cavity; under the reaction state, the filter element can prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the discharge state, the suction piece can suck the reaction cavity so as to enable the reagent to flow through the filter element to be discharged.

In one embodiment, the reagent discharging assembly is connected to the synthesis column, the reagent discharging assembly comprises a valve, a filter element is arranged in the reaction cavity, and the valve is arranged below the filter element; under the reaction state, the valve is closed to prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the discharge state, the valve is opened, so that the reagent flows through the filter element to be discharged.

In one embodiment, the DNA synthesis apparatus further comprises a turntable, the reagent input assembly comprises a plurality of reagent input members distributed along the circumferential direction, the synthesis columns are fixedly connected with the turntable, and the synthesis columns are driven by the turntable to rotate to the regions where the reagent input members are located.

In one embodiment, the DNA synthesizer includes a plurality of synthesis columns distributed along a circumferential direction, and each of the plurality of synthesis columns is fixedly connected to the turntable.

In one embodiment, the air inlet assembly further comprises an air inlet pipe, one end of the air inlet pipe is connected with the air source, the other end of the air inlet pipe penetrates through the central area of the rotary disc and then is connected with the air inlet part, and the air inlet part is located between the central area of the rotary disc and the synthesis column.

Above-mentioned DNA synthesizer, the inside reaction cavity that is formed with of synthetic post, can hold the nucleic acid synthesis carrier in this reaction cavity, and the required reagent of reaction is imported in the reaction cavity through reagent input assembly, reacts with the nucleic acid synthesis carrier, and after the reaction, the reagent discharges the reaction cavity through reagent discharge assembly. The gas source of the gas inlet assembly can provide gas, the gas enters the reaction cavity through the gas inlet piece, the gas oscillates and stirs the reagent in the reaction cavity, so that the nucleic acid synthesis carrier deposited at the bottom of the reagent is blown up and suspended in the reagent as much as possible, the area of the nucleic acid synthesis carrier, which can be in contact with the reagent, is increased, all binding sites on the surface of the nucleic acid synthesis carrier are in contact with the reagent and react as much as possible, and the synthesis rate is improved. In addition, the stirring position can be more accurate by the gas oscillation stirring mode.

The invention also provides a DNA synthesis method, which comprises the following steps:

s10, adding a nucleic acid synthesis carrier into the reaction cavity;

s20, adding a reagent into the reaction cavity;

s30, introducing gas into the reaction cavity to mix the nucleic acid synthesis carrier and the reagent in the reaction cavity in a shaking way;

s40, discharging the reagent out of the reaction cavity;

s50 repeats S20 to S40 a plurality of times.

In the DNA synthesis method, the gas oscillates and stirs the reagent in the reaction cavity, so that the nucleic acid synthesis carrier deposited at the bottom of the reagent is blown up and suspended in the reagent as much as possible, the area of the nucleic acid synthesis carrier which can be contacted with the reagent is increased, all binding sites on the surface of the nucleic acid synthesis carrier are contacted with the reagent and react as much as possible, and the synthesis rate is improved. In addition, the stirring position can be more accurate by the gas oscillation stirring mode.

Drawings

FIG. 1 is a schematic view of a partial structure of a DNA synthesizer according to an embodiment of the present invention;

FIG. 2 is a schematic diagram showing the structure of the turntable and the synthesis column of the DNA synthesis apparatus shown in FIG. 1;

FIG. 3 is a cross-sectional view of a synthesis column and a filter cartridge of the DNA synthesis apparatus of FIG. 1;

FIG. 4 is a sectional view of the synthesis column, the air inlet and the filter element of the DNA synthesis apparatus of FIG. 1.

Reference numerals:

a turntable 100;

a synthesis column 200, a main body 210, a reaction chamber 211, and an extension part 220;

a reagent input member 310, a reagent input valve 320, an input pipe 330;

the air inlet piece 410, the luer 411, the air inlet needle 412, the air inlet pipe 420, the air inlet valve 430, the flow meter 440 and the pressure regulating valve 450;

a filter element 510, a reagent discharge valve 520, and a discharge pipe 530.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the invention.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

In the present invention, unless otherwise expressly stated or limited, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through an intermediate. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.

It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like as used herein are for illustrative purposes only and do not denote a unique embodiment.

Referring to fig. 1 to 3, fig. 1 is a partial structural diagram of a DNA synthesizer according to an embodiment of the present invention, fig. 2 is a structural diagram of a turntable and a synthesis column of the DNA synthesizer of fig. 1, and fig. 3 is a sectional view of the synthesis column and a filter element of the DNA synthesizer of fig. 1. The DNA synthesizer according to an embodiment of the present invention includes a synthesis column 200, a reagent input unit, an air inlet unit, and a reagent discharge unit. The interior of the synthesis column 200 is formed with a reaction chamber 211, the reaction chamber 211 is used for accommodating nucleic acid synthesis carriers, and before reaction, an appropriate amount of nucleic acid synthesis carriers are placed in the reaction chamber 211. The reagent input assembly is used for inputting a reagent into the reaction cavity 211, and the reagent enters the reaction cavity 211 and then reacts with the nucleic acid synthesis carrier to be combined with the combination sites on the surface of the nucleic acid synthesis carrier. As is well known to those skilled in the art, oligonucleotide solid phase synthesis comprises four steps per cycle: deprotection, coupling, capping and oxidation, wherein different reagents need to be sequentially introduced into the synthesis column 200 in each cycle, and after the reaction of each reagent is finished, the reagent needs to be timely discharged from the reaction cavity 211, and nucleic acid with the expected length can be obtained on the nucleic acid synthesis carrier through multiple cycles. In this embodiment, the reagent discharging assembly discharges the reagent that finishes the reaction in the reaction chamber 211. The gas inlet assembly comprises a gas inlet 410 and a gas source for providing gas, the gas inlet 410 is connected with the synthesis column 200, and the gas provided by the gas source can flow through the gas inlet 410 into the reaction cavity 211. When the gas enters the reaction chamber 211, the nucleic acid synthesis carrier and the reagent in the reaction chamber 211 are agitated and stirred, and the nucleic acid synthesis carrier is blown up.

As is well known to those skilled in the art, the nucleic acid synthesis carrier is usually in the form of powder particles, and when the reagent enters the reaction chamber 211, the nucleic acid synthesis carrier is deposited on the bottom of the reagent, and a portion of the nucleic acid synthesis carrier accumulated inside may be difficult to sufficiently contact with the reagent and react, and the contact area between the portion of the nucleic acid synthesis carrier contacting the inner wall of the synthesis column 200 and the reagent is limited. In this embodiment, the gas oscillates and agitates the reagent in the reaction chamber 211, so that the nucleic acid synthesis carrier deposited at the bottom of the reagent is blown up and suspended in the reagent as much as possible, thereby increasing the area of the nucleic acid synthesis carrier that can contact the reagent, and allowing all binding sites on the surface of the nucleic acid synthesis carrier to contact the reagent and react as much as possible, thereby increasing the synthesis rate. In addition, the mode that the aeration body carries out the oscillation stirring can make the stirring position more accurate. The position of gas entering the reagent can be positioned in the area with lower synthesis rate through artificial design, so that the synthesis rate is greatly improved. Compared with the conventional methods such as mechanical stirring or magnetic stirring, the gas stirring method provided by the invention can realize micro regulation and control of the stirring degree and has higher precision.

Specifically, in some embodiments, the nucleic acid synthesis carrier may be a magnetic nucleic acid synthesis carrier, such as a magnetic bead coated with CPG, and the particle size of the magnetic nucleic acid synthesis carrier is preferably 0.5 μm to 100 μm, and may be, for example, 0.5 μm, 5 μm, 10 μm, 20 μm, 50 μm, 70 μm, or 100 μm. Of course, in other embodiments, the nucleic acid synthesis vector may be other types of substances that reversibly bind to nucleic acids.

The gas provided by the gas source needs to satisfy the conditions of stable chemical property, no reaction with the reagent, no reaction with the nucleic acid synthesis carrier, and the like. The gas is not particularly limited, and may be air, oxygen, an inert gas such as argon, or hydrogen. Specifically, in some embodiments, the gas may be nitrogen. Of course, in other embodiments, the gas may be an inert gas such as helium, argon, etc.

In some embodiments, the connection position of the air inlet 410 and the synthesis column 200 is located at one side of the reaction chamber 211, and the reagent input assembly is located above the reaction chamber 211. So set up and to make the two different positions that are located outside synthetic column 200 respectively of inlet piece 410 and reagent input assembly, the mounted position of the two can not take place to interfere to guarantee that reagent can add smoothly, and let in gas and carry out the oscillation. Specifically, the top of the synthesis column 200 is open, and the reagent input assembly adds the reagent from the top of the synthesis column 200 into the reaction chamber 211. When gas flows through the gas inlet 410 into the reaction chamber 211, the gas enters from one side of the reaction chamber 211 and flows out from the top of the synthesis column 200. Preferably, the connection location of the air inlet piece 410 and the synthetic column 200 is located at the upper end 1/5 ~ 1/2 of the synthetic column 200, for example 1/4.

Preferably, in some embodiments, the connection position of the air inlet 410 and the synthetic column 200 is located at one side of the reaction chamber 211, and the air inlet 410 is inclined upward. The upward inclination here means that the inlet end of the air induction member 410 is located outside the outlet end, and the inlet end is located above the outlet end, i.e., the connection position with the synthesis column 200. So configured, when gas flows through the gas inlet 410 into the reaction chamber 211, it may impact the reagent, which is inclined downward. Preferably, the axial angle of the air inlet 410 to the synthetic column 200 is 15 ° to 75 °, and may be 15 °, 30 °, 45 °, 60 °. Specifically, the direction of the gas impact on the reagent is from outside to inside and from top to bottom, which can cause a larger impact force, blow up as much nucleic acid synthesis carriers deposited at the bottom of the reagent as possible, and also form a small vortex at the bottom corner of the reaction chamber 211, reduce dead angles, so that the nucleic acid synthesis carriers in these regions can be blown up, thus further improving the synthesis rate.

Referring to FIGS. 3 and 4, FIG. 4 is a cross-sectional view of the synthesis column, the air inlet and the filter element of the DNA synthesizer of FIG. 1. Preferably, in some embodiments, the air intake 410 is inserted into the reaction chamber 211. The air outlet of the air inlet piece 410 can be positioned in the reagent by the arrangement, so that the oscillation stirring amplitude of the area where the air outlet is positioned is more violent, and the area with lower synthesis rate can be more accurately oscillated and stirred; in some embodiments, the gas outlet is located on the axis of the column 200 and is 1mm to 1cm from the upper surface of the filter element. After the gas enters the reagent, bubbles will escape, which escape from the top of the synthesis column 200. Preferably, in some embodiments, the air inlet 410 is inserted into the bottom of the reagent, so that the oscillating agitation amplitude can be further increased.

In some embodiments, the synthetic column 200 comprises a main body 210 and an extension 220, the extension 220 extends outwards from one side of the main body 210, the interior of the main body 210 forms a reaction cavity 211, the interior of the extension 220 is hollow and is communicated with the reaction cavity 211, and the air inlet 410 is connected with the extension 220. Specifically, the protruding portion 220 is inclined upward, i.e., the end of the protruding portion 220 connected to the air intake member 410 is located above the end of the protruding portion 220 connected to the main body portion 210. The air inlet 410 passes through the protrusion 220 into the reagent in the reaction chamber 211. As described above, this arrangement can cause a large impact force to blow up as much nucleic acid synthesis carriers deposited on the bottom of the reagent as possible, and can also form a small vortex at the bottom corner of the reaction chamber 211 to reduce dead corners and blow up nucleic acid synthesis carriers in these areas, thereby further improving the synthesis rate. Preferably, in some embodiments, the radial dimension of main body portion 210 gradually decreases in a vertically downward direction, that is, main body portion 210 is funnel-shaped, and the upper dimension of main body portion 210 is larger, so as to facilitate adding reagents into reaction cavity 211. Preferably, in some embodiments, the main body 210 and the extension 220 are integrally formed as a single body, so that no additional sealing structure is required to seal therebetween, and the structure is simple. Preferably, in some embodiments, the main body portion 210 and the extension portion 220 may be integrally formed by injection molding.

In some embodiments, the gas inlet 410 extends through the extension 220 to a radially central position of the reaction chamber 211. Therefore, the air outlet of the air inlet member 410 is located in the center area of the reagent in the radial direction, so that the area around the air outlet is located in the oscillating and stirring range as much as possible, the probability of dead angles is reduced, the nucleic acid synthesis carrier can be blown up as much as possible, and the synthesis rate is higher.

In some embodiments, the main body 210 is connected to a plurality of protruding portions 220, and the gas can be introduced into different regions of the reaction chamber 211 through the plurality of protruding portions 220 for oscillating and stirring, so as to increase the stirring amplitude and further improve the synthesis rate. Preferably, in some embodiments, the plurality of protruding portions 220 are uniformly distributed on the sidewall of the main body portion 210 along the circumferential direction, each protruding portion 220 has one air inlet 410, and the connection line of the positions of the plurality of air inlet 410 protruding into the reagent is annular. So set up can be comparatively even each regional oscillation stirring in to reagent.

Referring to fig. 1, 3 and 4, in some embodiments, the air inlet 410 includes a male luer 411 and an air inlet needle 412, the male luer 411 is fixedly connected with the air inlet needle 412, and the air inlet needle 412 is inserted into the protruding portion 220 and is in interference fit with the protruding portion 220. Specifically, the luer 411 is connected to the gas inlet tube 420, and the gas enters the luer 411 through the gas inlet tube 420, and then flows through the gas inlet needle 412 into the reaction chamber 211. As mentioned above, the synthetic column 200 can be manufactured by injection molding, and the synthetic column 200 has a certain micro elasticity, so that the air inlet 410 can be clamped in the extension part 220, and during installation, the air inlet 410 only needs to be plugged into the extension part 220, and the assembly is simple. And luer 411 is standard, and direct purchase can, need not special manufacture. Preferably, the air inlet needle 412 is made of teflon with stable chemical property and good corrosion resistance, so as to ensure that the air inlet needle 412 is not corroded by the reagent and does not pollute the reagent.

In some embodiments, a reagent discharge assembly is coupled to the synthesis column 200, the reagent discharge assembly comprising a suction member, a filter element 510 disposed within the reaction chamber 211; in the reaction state, the filter element 510 can prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the discharge state, the suction member can suck the reaction chamber 211 so as to discharge the reagent flowing through the filter element 510. Specifically, the filter element 510 has a pore size in the range of 0.4 μm to 0.6 μm, and the filter element 510 is hydrophobic and permeable to air. Filter element 510 is located in a bottom end region of body portion 210 and is in interference fit with an inner wall of body portion 210. Because the filter element 510 has hydrophobicity and small pores, in the reaction state, when no external force is applied, the filter element 510 can seal the bottom end of the reaction cavity 211, so that the reagent and the nucleic acid synthesis carrier are sealed in the reaction cavity 211 for reaction. After the reaction is completed, in the discharge state, the reaction chamber 211 is sucked from the lower side of the filter element 510 by the suction member, and the reagent in the reaction chamber 211 flows downward by the negative pressure adsorption formed during the suction, and flows through the pores of the filter element 510 to be discharged out of the reaction chamber 211. The size of the nucleic acid synthesis carrier is larger than the pore size of the filter element 510, so that the nucleic acid synthesis carrier can still remain in the reaction chamber 211 and be blocked by the filter element 510 when the suction piece performs negative pressure adsorption. In a specific embodiment, PPFE with a diameter of 3.8mm and a thickness of 3.3mm is selected as the filter element 510. Specifically, the bottom end of the synthesis column 200 is connected with a discharge pipe 530, the discharge pipe 530 is connected with a suction member, and a reagent discharge valve 520 is provided on the discharge pipe 530. Opening the reagent discharge valve 520 opens the discharge pipe 530, and opening the suction member draws the reaction chamber 211 from below the filter element 510 through the discharge pipe 530, thereby discharging the reagent through the discharge pipe 530. Specifically, the reagent exhaust valve 520 may be a solenoid valve. In the above embodiment, the blocking and discharging of the reagent are realized by the filter element 510, so that the structure is simple, the operation is very convenient, and when the reagent needs to be discharged, only the reagent discharge valve 520 and the suction piece need to be opened.

In other embodiments, a reagent discharge assembly is connected to the synthesis column 200, the reagent discharge assembly comprises a valve, a filter element 510 is disposed in the reaction chamber 211, and the valve is disposed below the filter element 510; in the reaction state, the valve is closed to prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the drain state, the valve is opened to allow reagent to flow through the cartridge 510 for draining. Specifically, filter element 510 is located at a bottom end region of body portion 210 and is in interference fit with an inner wall of body portion 210. In the reaction state, the valve is closed, and the bottom end of the reaction cavity 211 can be closed, so that the reagent and the nucleic acid synthesis carrier are sealed in the reaction cavity 211 for reaction. When the valve is opened in the discharge state after the reaction is completed, the reagent will flow through the pores of the filter element 510 and be discharged out of the reaction chamber 211. The size of the nucleic acid synthesis vector is larger than the size of the pores of the filter element 510, and thus, the nucleic acid synthesis vector is blocked by the filter element 510 above the filter element 510. In this embodiment, the filter element 510 is not made of hydrophobic material, and the reagent can flow through the filter element 510 when no external force is applied. In this embodiment, some of the reagent may flow through the filter element 510 to the gap between the filter element 510 and the valve. Preferably, the top surface of the valve is brought into contact with the bottom surface of the filter element 510, that is, the gap between the filter element 510 and the valve is minimized, so that the reagent is located above the filter element 510 as much as possible to react with the nucleic acid synthesis carrier, thereby increasing the synthesis rate.

Referring to fig. 1, in some embodiments, the DNA synthesis apparatus further comprises a carousel 100, and the reagent input assembly comprises a plurality of reagent inputs 310, the plurality of reagent inputs 310 being distributed along a circumferential direction of the carousel 100. The synthesis column 200 is fixedly connected to the turntable 100, and the synthesis column 200 is driven by the turntable 100 to rotate to the area where each reagent input member 310 is located. Specifically, the synthesis column 200 is fixed on the top of the turntable 100, the reagent input member 310 is connected to the reagent bottle through the input pipe 330, the reagent input valve 320 is disposed on the input pipe 330, and the connection and disconnection of the input pipe 330 can be realized by adjusting the reagent input valve 320. When the turntable 100 rotates, the synthesis column 200 will be driven to rotate synchronously, and thus reach the lower part of each reagent input member 310 in turn. When the synthesis column 200 is rotated to a position below a certain reagent input member 310, the reagent input member 310 may add a corresponding reagent to the synthesis column 200. When the reaction in this step is completed, the reagents are discharged in the above manner, and then the turntable 100 is rotated to transport the synthesis column 200 to the position below the reagent input member 310 corresponding to the reaction in the next step. By the arrangement, each reaction step can be completed conveniently and quickly. Preferably, the synthesis column 200 is located at a region close to the edge of the turntable 100, and the radius of the circle formed by the reagent inlets 310 can be larger, the circumferential distance between the reagent inlets 310 can be larger, the arrangement is not too crowded, and the reagent inlets do not easily interfere with each other.

Further, in some embodiments, the DNA synthesizer includes a plurality of synthesis columns 200 distributed along the circumferential direction, and each of the plurality of synthesis columns 200 is fixedly connected to the turntable 100. The radius of the circle enclosed by the plurality of synthesis columns 200 is equal to the radius of the circle enclosed by the plurality of reagent inputs 310. When the turntable 100 rotates, the plurality of synthesis columns 200 will be driven to rotate synchronously, and the reagents will reach the lower part of the corresponding reagent input member 310 respectively for reagent addition. The angle of each rotation of the turntable 100 is the same as the angle of the adjacent combining post 200. For example, when the turntable 100 is rotated by a certain angle, the first synthesis column 200 reaches below the first reagent input member 310, and the first reagent input member 310 adds the first reagent to the first synthesis column 200 and performs a corresponding reaction. When the turntable 100 continues to rotate by a certain angle, the first synthesis column 200 reaches the position below the second reagent input part 310, and the second reagent input part 310 adds the second reagent into the first synthesis column 200 and carries out corresponding reaction; at the same time, the second synthesis column 200 adjacent to the first synthesis column 200 reaches below the first reagent input member 310, and the first reagent input member 310 adds the first reagent to the second synthesis column 200 and performs a corresponding reaction. Through the arrangement, the assembly line type operation can be realized, so that the plurality of synthesis columns 200 can synchronously carry out respective reaction, and the efficiency can be greatly improved.

In some embodiments, the air inlet pipe 420 is connected to the air source at one end and connected to the air inlet 410 after passing through the central region of the turntable 100, and the air inlet 410 is located between the central region of the turntable 100 and the synthesis column 200. The air inlet piece 410 can not occupy the position outside the radial range of the turntable 100, so that the component arrangement is more compact and the space is more saved. Specifically, the top end of the air inlet pipe 420 passes through the central region of the turntable 100 and then is connected with the air inlet member 410, the bottom end is connected with the air source, and the air inlet pipe 420 is provided with an air inlet valve 430, a flow meter 440, a pressure regulating valve 450 and the like. The pressure regulating valve 450 can regulate the gas pressure output by the gas source to obtain a proper oscillation stirring amplitude. By adjusting the parameters of the flow meter 440, the gas can be made to flow at a more appropriate rate. When the gas inlet 410 protrudes into the reagent, the flow rate can preferably be adjusted so that the gas forms small bubbles that are continuous as it enters the reagent. When the reaction is completed without agitation, the gas inlet valve 430 may be closed to stop the gas input.

In some embodiments, the present invention also provides a DNA synthesis method comprising the steps of:

s10, adding a nucleic acid synthesis carrier into the reaction cavity;

s20, adding a reagent into the reaction cavity;

s40, discharging the reagent out of the reaction cavity;

s50 repeats S20 to S40 a plurality of times. In repetition, different reagents may be added each time.

In the DNA synthesis method, gas oscillates and stirs the reagent in the reaction cavity 211, so that the nucleic acid synthesis carrier deposited at the bottom of the reagent is blown up and suspended in the reagent as much as possible, thereby increasing the area of the nucleic acid synthesis carrier which can be contacted with the reagent, and enabling all binding sites on the surface of the nucleic acid synthesis carrier to be contacted with the reagent and react as much as possible, thereby improving the synthesis rate. In addition, the stirring position can be more accurate by the gas oscillation stirring mode.

In the stirring process of mechanical stirring and magnetic stirring, a stirring blind area may exist, and the invention drives the all-dimensional liquid flow in the reaction cavity 211 of the synthesis column 200 by adjusting the gas introduction angle, the distance between the position of the gas entering the reagent and the filter element 510 and the gas flow rate, so that the mixture of the carrier and the reagent is more uniform, the uniformity of the product is improved, and the yield is further improved.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. DNA synthesis device, is characterized in that, comprises:

a synthesis column, wherein a reaction cavity is formed inside the synthesis column, and the reaction cavity is used for accommodating a nucleic acid synthesis carrier;

a reagent input component, the reagent input component is used for inputting a reagent into the reaction chamber;

an air intake assembly, the air intake assembly includes an air intake part and an air source, the air source is used for providing gas, the air intake part is connected with the synthesis column, and the gas enters the a reaction chamber to oscillate the nucleic acid synthesis carrier and reagents in the reaction chamber;

The reagent discharge component is used for discharging the reagent that has finished the reaction in the reaction chamber.

2 . The DNA synthesis device according to claim 1 , wherein the connection position of the gas inlet member and the synthesis column is located at one side of the reaction chamber, and the reagent input assembly is located in the reaction chamber. 3 . above the body.

3 . The DNA synthesis device according to claim 2 , wherein the synthesis column comprises a main body part and a protruding part, the protruding part protrudes outward from one side of the main body part, and the main body The reaction cavity is formed in the interior of the part, the interior of the protruding part is hollow and communicated with the reaction cavity, and the air inlet is connected with the protruding part.

4 . The DNA synthesis apparatus according to claim 3 , wherein the air inlet member extends into the center of the reaction chamber along the radial direction through the protruding portion. 5 .

5 . The DNA synthesis device according to claim 3 , wherein the air inlet member comprises a luer connector and an air intake needle, the luer connector is fixedly connected to the air intake needle, and the air intake needle is fixedly connected. 6 . Insert the protrusions with an interference fit.

6 . The DNA synthesis device according to claim 1 , wherein the reagent discharge assembly is connected to the synthesis column, the reagent discharge assembly comprises a suction part, and a filter element is arranged in the reaction chamber; the reaction state In the discharge state, the filter element can prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the discharge state, the suction member can suction the reaction chamber, so that the reagent flows through the filter element and is discharged.

7. The DNA synthesis device according to claim 1, wherein the reagent discharge assembly is connected to the synthesis column, the reagent discharge assembly comprises a valve, a filter element is provided in the reaction chamber, and the valve is provided with a valve. Under the filter element; in the reaction state, the valve is closed to prevent the reagent and the nucleic acid synthesis carrier from flowing out; in the discharge state, the valve is opened to allow the reagent to flow through the filter element to be discharged.

8 . The DNA synthesis device according to claim 1 , wherein the DNA synthesis device further comprises a turntable, the reagent input assembly comprises a plurality of reagent input members distributed in a circumferential direction, and the synthesis column is connected to the The turntable is fixedly connected, and the synthesis column is driven by the turntable to rotate to the area where each of the reagent input pieces is located.

9 . The DNA synthesis device according to claim 8 , wherein the DNA synthesis device comprises a plurality of the synthesis columns distributed along the circumferential direction, and the plurality of the synthesis columns are all fixedly connected to the turntable. 10 .

10. DNA synthesis method, is characterized in that, comprises the steps:

S10 adds a nucleic acid synthesis carrier into the reaction chamber;

S20 adding reagents into the reaction chamber;

S30, gas is introduced into the reaction chamber to shake and mix the nucleic acid synthesis carrier and the reagent in the reaction chamber;

S40, discharging the reagent out of the reaction chamber;

S50 repeats S20 to S40 multiple times.