CN215209698U

CN215209698U - Automatic library construction device based on closed card box

Info

Publication number: CN215209698U
Application number: CN202023259631.5U
Authority: CN
Inventors: 李松; 潘文京; 汤从利; 刘洪娜
Original assignee: Nanjing Arix Biotechnology Co ltd
Current assignee: Nanjing Arix Biotechnology Co ltd
Priority date: 2020-12-29
Filing date: 2020-12-29
Publication date: 2021-12-17
Anticipated expiration: 2030-12-29

Abstract

The utility model provides an automatic library founds device based on closed card box, including the card box main part, the inboard bottom center of card box main part is provided with the base, seted up a waste liquid storage tank, one by left to right on the base and built storehouse reaction tank and thirteen reagent and deposit the pond, install the guide rail on the inner wall that card box main part is parallel with the base, install the slider in the guide rail, be provided with the arm on the slider, install the pipetting gun on the arm, pipetting gun and arm all are connected at the outside controller of card box main part through pipe and setting, install the cylinder on the outer wall of card box main part, practice thrift manpower and materials, avoid the testing result to be influenced by the operating factor, effectually avoid polluting.

Description

Automatic library construction device based on closed card box

Technical Field

The utility model relates to a gene detection technology field especially relates to an automatic library founds device based on closed card box.

Background

With the rapid development of the field of biotechnology, the molecular level is deeply explored to obtain more accurate individual nucleic acid information, which is a necessary condition for improving the human life health quality and preventing diseases. The cDNA library has special advantages in the research of the expression state of genome and the function identification of expressed gene in a specific cell, so that the cDNA library has wider application value in the research of life phenomena such as ontogenesis, cell differentiation, cell cycle regulation, cell senescence, death regulation and the like, and is the most commonly used gene library in research work. Thus, the construction of libraries is a key step in obtaining individualized nucleic acid information.

Conventional library building steps are complicated, after the extracted and purified genome DNA is obtained, the genome DNA is processed according to the research purpose, the DNA is randomly sheared into fragments with different lengths, the tail end is repaired, or the genome DNA is digested by restriction endonuclease, the DNA fragment with proper length is found, the target DNA molecule is obtained by purification from the solution, a joint, a signal recognition molecule and the like are added for PCR amplification reaction, the library is enriched after the amplification reaction is finished, the library concentration is determined, a standardized sample is built, all samples are mixed, and then the computer is used for sequencing. The whole library building process needs manual operation for 6-11 hours, generally takes 2-3 days, and is time-consuming and labor-consuming.

Meanwhile, the library construction experiment has high requirements on sites and operators. The experiment site needs to be strictly partitioned, and the operation of corresponding steps is carried out in different partitioned areas, so that the experiment is prevented from being carried out alternately, and the pollution is reduced. The operator who builds the storehouse needs to possess corresponding professional knowledge and abundant experience, and the skilled experiment that carries out guarantees the stability of result. However, human operation is different from a machine, and all the flow paths of each operation cannot be guaranteed to be completely consistent, so that uncontrollable performance exists. Therefore, the detection result is affected by the operation factor, and the contamination cannot be completely avoided.

Although contamination is minimized in all respects, there is a possibility of contamination as long as the reaction tube is uncovered. In order to shorten the reaction time, avoid the pollution caused by repeated uncapping and reduce manual operation, we propose automated library construction based on closed cartridges.

SUMMERY OF THE UTILITY MODEL

The utility model aims at solving the defects existing in the prior art, and providing an automatic library constructing device which can be automatically carried out in a closed environment, does not need additional operation and greatly saves manpower.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

design an automatic library construction device based on closed card box, including the card box main part, its characterized in that: the card box comprises a card box body and is characterized in that a base is arranged at the center of the bottom of the inner side of the card box body, a waste liquid storage pool, a library building reaction pool and thirteen reagent storage pools are arranged on the base from left to right, a guide rail is arranged on an inner wall of the card box body parallel to the base, a sliding block is arranged in the guide rail, a mechanical arm is arranged on the sliding block, a liquid transfer gun is arranged on the mechanical arm, the liquid transfer gun and the mechanical arm are both connected with a controller arranged outside the card box body through a guide pipe, and an air cylinder is arranged on the outer wall of the card box body.

Preferably, the thirteen reagent storage pools are a reagent storage pool 1, a reagent storage pool 2, a reagent storage pool 3, a reagent storage pool 4, a reagent storage pool 5, a reagent storage pool 6, a reagent storage pool 7, a reagent storage pool 8, a reagent storage pool 9, a reagent storage pool 10, a reagent storage pool 11, a reagent storage pool 12 and a reagent storage pool 13.

Preferably, 400 μ L of O sealing liquid, 1500 μ L of buffer solution, 2500 μ L of buffer solution, 500 μ L of ethanol, 170 μ L of magnetic particles, 3200 μ L of buffer solution, 4500 μ L of buffer solution, 1200 μ L of amplification solution, 220 μ L of amplification solution, 5500 μ L of buffer solution, 3200 μ L of amplification solution are sequentially placed in the reagent storage pool 1, the reagent storage pool 2, the reagent storage pool 3, the reagent storage pool 4, the reagent storage pool 5, the reagent storage pool 6, the reagent storage pool 7, the reagent storage pool 8, the reagent storage pool 9, the reagent storage pool 10, and the reagent storage pool 11, and the reagent storage pool 13(15) is 2200 μ L of magnetic particles.

The utility model provides an automatic library founds device based on closed card box, beneficial effect lies in: the library construction is automated throughout the process in a closed environment. The library is built through the card box, and various problems in the prior art are solved. When the library is built, only a nucleic acid sample needs to be added into the reaction hole, no additional operation is needed, and the labor is greatly saved. Meanwhile, the reaction is carried out in the closed card box, the cover does not need to be opened in the whole process, pollution hardly exists, a plurality of fields do not need to be provided for experiments, the warehouse building process is mechanical, the influence of human factors is avoided, the whole warehouse building reaction only needs about 10 hours, and the reaction time is greatly shortened.

Drawings

Fig. 1 is a schematic structural diagram of the present invention;

fig. 2 is a schematic view of the base structure provided by the present invention.

In the figure: 1 waste liquid storage tank, 2 build storehouse reaction tank, 3 the

reagent deposit

1, 4 the

reagent deposit

2, 5 the

reagent deposit

3, 6 the

reagent deposit

4, 7 the

reagent deposit

5, 8 the

reagent deposit

6, 9 the reagent deposit 7, 10 the

reagent deposit

8, 11 the reagent deposit 9, 12 the reagent deposit 10, 13 the reagent deposit 11, 14 the reagent deposit 12, 15 the

reagent deposit

13, 16 locating holes, 17 sliders, 18 bases, 19 liquid-transfering guns, 20 cylinders, 21 pipes, 22 arms, 23 guide rails, 24 card box main parts.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments.

Referring to fig. 1-2, the utility model provides an automatic library founds device based on closed card box, including card box main part 24, card box main part 24's inboard bottom center is provided with base 18, a waste liquid storage tank 1, one build storehouse reaction tank 2 and thirteen reagent storage tanks have been seted up from left to right on the base 18, install guide rail 23 on card box main part 24 and an inner wall that base 18 is parallel, install slider 17 in the guide rail 23, be provided with arm 22 on the slider 17, install pipetting gun 19 on the arm 22, pipetting gun 19 and arm 22 all are connected with the controller that sets up at card box main part 24 outside through the pipe, install cylinder 20 on card box main part 24's the outer wall.

Thirteen reagent storage pools are a reagent storage pool 1, a reagent storage pool 2, a reagent storage pool 3, a reagent storage pool 4, a reagent storage pool 5, a reagent storage pool 6, a reagent storage pool 7, a reagent storage pool 8, a reagent storage pool 9, a reagent storage pool 10, a reagent storage pool 11, a reagent storage pool 12 and a reagent storage pool 13, wherein the reagent storage pool 13, the reagent storage pool 2, the reagent storage pool 3, the reagent storage pool 4, the reagent storage pool 5, the reagent storage pool 6, the reagent storage pool 7, the reagent storage pool 8, the reagent storage pool 9, the reagent storage pool 10 and the reagent storage pool 11 are sequentially filled with 400 muL of sealing liquid, 1500 muL of buffer liquid, 2500 muL of buffer liquid, 500 muL of ethanol, 170 muL of magnetic particles, 3200 muL of buffer liquid, 4500 muL of buffer liquid, 1200 muL of amplification liquid, 220 muL of amplification liquid and 5500 muL of buffer liquid, 3200. mu.L of amplification solution, 2200. mu.L of magnetic particles in the reagent storage pool 13 (15).

The waste liquid hole is used for storing waste container liquid after reaction in each step is finished, the reaction hole is closed in the card box after nucleic acid is added through opening a cover in the first step, multi-step reaction is carried out to complete library construction, the reagent storage hole stores reagents required by each step of reaction, and the reagents are transferred to the reaction hole in sequence by using a mechanical arm in the card box to carry out experiments. The mechanical arm replaces manual storehouse building of operating personnel, only needs the first step to add the nucleic acid sample in the reaction hole, and the operation degree of difficulty greatly reduced only needs the personnel that have the experiment basis to operate, and labour saving and time saving has avoided manual operation's uncontrollable nature, guarantees that the experiment is stable to be gone on, falls to minimum with manual operation's influence. Except adding the sample, all reactions are carried out in a closed environment, so that the pollution possibly caused by opening the cover for many times is avoided, the strict zoning operation on the field is not needed, and the pollution is minimized.

Fully vibrating the card box before use, adding the extracted paper blood nucleic acid into the No. 2 reaction hole by using a pipette, covering a cover to seal the card box, sending the card box into an automatic library building system instrument, selecting a paper blood library building program on a computer and running, and reacting for about 10 hours to complete automatic library building.

The above, only be the concrete implementation of the preferred embodiment of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art is in the technical scope of the present invention, according to the technical solution of the present invention and the utility model, the concept of which is equivalent to replace or change, should be covered within the protection scope of the present invention.

Claims

1. An automated library building apparatus based on closed cassettes, comprising a cassette body (24), characterized in that: the bottom center of the inner side of the card box main body (24) is provided with a base (18), the base (18) is provided with a waste liquid storage pool (1), a warehouse building reaction pool (2) and thirteen reagent storage pools from left to right, a guide rail (23) is arranged on an inner wall of the card box main body (24) parallel to the base (18), a sliding block (17) is arranged in the guide rail (23), a mechanical arm (22) is arranged on the sliding block (17), a liquid transferring gun (19) is arranged on the mechanical arm (22), the liquid transferring gun (19) and the mechanical arm (22) are both connected with a controller arranged outside the card box main body (24) through a guide pipe, and an air cylinder (20) is arranged on the outer wall of the card box main body (24).

2. The closed cassette-based automated library building apparatus of claim 1, wherein: thirteen reagent storage pools are a reagent storage pool 1(3), a reagent storage pool 2(4), a reagent storage pool 3(5), a reagent storage pool 4(6), a reagent storage pool 5(7), a reagent storage pool 6(8), a reagent storage pool 7(9), a reagent storage pool 8(10), a reagent storage pool 9(11), a reagent storage pool 10(12), a reagent storage pool 11(13), a reagent storage pool 12(14) and a reagent storage pool 13 (15).

3. The closed cassette-based automated library building apparatus of claim 1, wherein: the reagent storage pool 1(3), the reagent storage pool 2(4), the reagent storage pool 3(5), the reagent storage pool 4(6), the reagent storage pool 5(7), the reagent storage pool 6(8), the reagent storage pool 7(9), the reagent storage pool 8(10), the reagent storage pool 9(11), the reagent storage pool 10(12) and the reagent storage pool 11(13) are sequentially provided with sealing liquid 400 mu L, buffer solution 1500 mu L, buffer solution 2500 mu L, ethanol 500 mu L, magnetic particles 170 mu L, buffer solution 3200 mu L, buffer solution 4500 mu L, amplification solution 1200 mu L, amplification solution 220 mu L, buffer solution 5500 mu L and amplification solution 3200 mu L, and the reagent storage pool 13(15) is magnetic particles 2200 mu L.