Nucleic acid extraction kit

Abstract

The nucleic acid extraction kit comprises a liquid inlet and outlet pipe and a suction pipe cavity, the suction pipe cavity is made of rubber materials and communicated with the top end of the liquid inlet and outlet pipe, a switch valve is arranged on the liquid inlet and outlet pipe, and a lysis solution bin, a magnetic bead liquid bin, a washing liquid bin and an eluent bin are arranged on the suction pipe cavity respectively. Pricking needle assemblies are respectively arranged on separation surfaces of the lysate bin, the magnetic bead liquid bin, the washing liquid bin and the eluent bin and the suction tube cavity, a magnetic sheet slot is further formed in the outer wall surface of the suction tube cavity, and a magnetic sheet can be inserted into the magnetic sheet slot. Reagents required by extraction are pre-stored in the corresponding reagent bin integrated with the suction tube cavity, so that the portable nucleic acid extraction device is convenient to carry outdoors for nucleic acid extraction operation; in the extraction process, reagents can be rapidly added into the suction tube cavity by extruding the pricking needle assembly to pierce the corresponding reagent bin, a pipette is not needed, external magnetic field force can be provided by inserting or taking out the magnetic sheet into or from the magnetic sheet slot, operation is simple and rapid, and the extraction efficiency is effectively improved.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of biotechnology, concretely relates to a nucleic acid extraction kit. BACKGROUND

[0002] The magnetic bead method for extracting nucleic acid refers to the preparation of superparamagnetic silica nano magnetic beads after the surface modification and surface modification of superparamagnetic nanoparticles, and the nucleic acid molecules can be specifically recognized and efficiently combined with the magnetic beads due to the rich surface active groups of the magnetic beads, and the separation from the sample to be tested can be realized under the action of the external magnetic field. The magnetic bead method for extracting nucleic acid generally includes four steps of lysis, combination, washing and elution. In the prior art, small batches of nucleic acid are usually extracted manually. The lysis, combination and rinsing steps in the extraction process are carried out in different centrifugal tubes. The corresponding reagents are sucked into the centrifugal tubes by a handheld pipette with a gun head. The reagent adding operation steps are complex and tedious, time-consuming and laborious. If not careful, liquid may drop outside, affecting the extraction efficiency. Moreover, it is not convenient to carry out extraction operations outdoors. SUMMARY

[0003] The main purpose of the utility model is to overcome the defects in the background art, and provide a nucleic acid extraction kit.

[0004] To achieve the above-mentioned purpose, the nucleic acid extraction kit provided by the utility model comprises an inlet and outlet pipe and a straw cavity. The straw cavity is made of rubber material. The straw cavity is in communication with the top end of the inlet and outlet pipe. A switch valve is arranged on the inlet and outlet pipe. A lysis liquid bin, a magnetic bead liquid bin, a washing liquid bin and an elution liquid bin are arranged on the straw cavity respectively. A needle assembly is arranged on the separation surface of the lysis liquid bin, the magnetic bead liquid bin, the washing liquid bin and the elution liquid bin respectively. A magnetic sheet slot is further arranged on the outer wall surface of the straw cavity. The magnetic sheet slot can insert a magnetic sheet. The reagents required for the lysis, combination, washing and elution steps of the nucleic acid sample are stored in the corresponding reagent bins integrated with the straw cavity. It is convenient to carry out nucleic acid extraction outdoors. In the extraction process, the reagents pre-stored in the reagent bins are squeezed into the straw cavity after the corresponding reagent bins and the straw cavity are connected by the needle assembly. The corresponding reagents can be added for the corresponding extraction steps. Since the reagents do not need to be transferred by a pipette, the operation is simple and fast. In the extraction process, the magnetic sheet is inserted into the magnetic sheet slot or taken out to provide an external magnetic field to adsorb and release the magnetic beads. The convenience of the extraction operation is effectively improved. The nucleic acid extraction process is accelerated and the extraction efficiency is improved.

[0005] Further optimize the technical scheme. The inlet and outlet pipe is provided with a scale line. The scale line is convenient for controlling the suction amount of the nucleic acid sample.

[0006] Further optimization technical scheme, the material of the inlet and outlet pipe is plastic or glass.

[0007] Further optimization technical scheme, the inner wall of the straw cavity is provided with a corrosion-resistant layer, and the corrosion-resistant layer is composed of a polytetrafluoroethylene coating.

[0008] Further optimization technical scheme, the switch valve comprises a valve rod and a ball, the ball is arranged in the interior of the inlet and outlet pipe, the valve rod is rotatable and penetrates into the interior of the inlet and outlet pipe and is connected with the ball, and one end of the valve rod located outside the inlet and outlet pipe is provided with a rotating handle.

[0009] Further optimization technical scheme, the top of the straw cavity is provided with a gas vent, and the gas vent is provided with a plug.

[0010] Further optimization technical scheme, the piercing needle assembly is fixedly arranged on the bottom surfaces of the lysis liquid bin, the magnetic bead liquid bin, the washing liquid bin and the elution liquid bin respectively; the piercing needle assembly comprises a U-shaped elastic sheet and a puncture needle, the lower wall of the U-shaped elastic sheet is fixedly connected with the bottom surfaces of the lysis liquid bin, the magnetic bead liquid bin, the washing liquid bin and the elution liquid bin after being turned over by 90 degrees, a puncture hole is formed in the lower wall of the U-shaped elastic sheet, the puncture needle is arranged in the opening of the U-shaped elastic sheet and is fixedly connected with the upper side wall of the U-shaped elastic sheet, and the puncture needle corresponds to the puncture hole.

[0011] Further optimization technical scheme, the nucleic acid extraction kit further comprises a PCR tube corresponding to the bottom end of the inlet and outlet pipe, used for collecting the extracted nucleic acid extraction liquid.

[0012] The nucleic acid extraction kit has the advantages that the reagents required in the lysis, combination and rinsing steps are stored in the corresponding reagent bins in advance and integrated with the straw cavity, so that the nucleic acid extraction operation can be conveniently carried out outdoors; in the extraction process, after the separating surface between the corresponding reagent bin and the straw cavity is pierced by the piercing needle assembly, the corresponding reagent bin can be squeezed to quickly add the reagent into the straw cavity, without using a pipette, and the operation is simple and fast; by inserting or taking out the magnetic sheet into or from the magnetic sheet slot, an external magnetic field force is provided in the processes of releasing nucleic acid from microorganisms, adsorbing nucleic acid by magnetic beads, removing impurities by washing and eluting nucleic acid, and the extraction efficiency is effectively improved. BRIEF DESCRIPTION OF DRAWINGS

[0013] Fig. 1It is the whole schematic view of the nucleic acid extraction kit in the embodiment of the utility model.

[0014] Fig. 2 It is the schematic view of the switch valve in the embodiment of the utility model.

[0015] Fig. 3 It is the schematic view of the switch valve in the embodiment of the utility model.

[0016] Fig. 4 It is the schematic view of the needle assembly in the embodiment of the utility model.

[0017] Reference signs: 1 inlet and outlet pipe; 2 straw cavity; 201 anticorrosive layer; 202 air release port; 203 plug cap; 3 switch valve; 301 valve rod; 302 ball; 303 rotating handle; 4 lysis liquid bin; 5 magnetic bead liquid bin; 6 washing liquid bin; 7 elution liquid bin; 8 needle assembly; 801 U-shaped elastic sheet; 802 puncture needle; 803 puncture hole; 9 magnetic sheet slot; 10 magnetic sheet; 11 scale line; 12 PCR tube. DETAILED DESCRIPTION

[0018] In order to make the technical problems, technical schemes and beneficial effects to be solved in the embodiments of the utility model more clearly understood, the utility model will be further described in detail below in combination with the drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the utility model, and are not used to limit the utility model.

[0019] It should be noted that when an element is referred to as being "fixed to" or "disposed on" another element, it can be directly on the other element or indirectly on the other element. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or indirectly connected to the other element. In addition, the connection can be for fixing or for circuit communication.

[0020] It should be understood that the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only used to facilitate the description of the embodiments of the utility model and simplify the description, and therefore cannot be understood as indicating or implying that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the utility model.

[0021] In addition, the terms "first", "second", "third", "fourth", "fifth", "sixth", "seventh" and "eighth" are only used for descriptive purpose and are not to be construed as indicating or implying relative importance or a specific number of the indicated technical features. Thus, features defined with "first", "second", "third", "fourth", "fifth", "sixth", "seventh" and "eighth" can explicitly or implicitly include one or more of the features. In the description of the embodiments of the present application, the meaning of "a plurality of" is two or more, unless otherwise explicitly and specifically limited.

[0022] Please refer to Figs. 1 to 4The nucleic acid extraction kit disclosed in an embodiment comprises an inlet and outlet pipe 1 and a straw cavity 2 connected with the top end of the inlet and outlet pipe 1, the inlet and outlet pipe 1 is made of plastic or glass material, the straw cavity 2 is made of elastic rubber material, and an anticorrosive layer 201 is arranged on the inner wall surface of the straw cavity 2, the anticorrosive layer 201 is composed of a polytetrafluoroethylene coating, the straw cavity 2 has a gasbag structure and can generate a gas pressure difference after being pressed to suck liquid into the inlet and outlet pipe 1 and / or the straw cavity 2, a switch valve 3 is arranged on the inlet and outlet pipe 1 and located at the top of the inlet and outlet pipe 1, the switch valve 3 is closed after the liquid is sucked into the straw cavity 2 to prevent the liquid from flowing out, and a lysis liquid bin 4, a magnetic bead liquid bin 5, a washing liquid bin 6 and an elution liquid bin 7 are arranged on the straw cavity 2 respectively, specifically, the lysis liquid bin 4, the magnetic bead liquid bin 5, the washing liquid bin 6 and the elution liquid bin 7 are used to store reagents required for lysis, combination, washing and elution steps of the nucleic acid sample, the lysis liquid bin 4, the magnetic bead liquid bin 5, the washing liquid bin 6 and the elution liquid bin 7 can be arranged around one-half of the outer wall surface area of the straw cavity 2, and the other half of the area is reserved for pressing operation, of course, the lysis liquid bin 4, the magnetic bead liquid bin 5, the washing liquid bin 6 and the elution liquid bin 7 can also be arranged separately in the interior of the straw cavity 2, as long as at least one of the separating surfaces of the lysis liquid bin 4, the magnetic bead liquid bin 5, the washing liquid bin 6 and the elution liquid bin 7 is attached to the wall surface of the straw cavity 2 or shared, a needle assembly 8 is arranged on the separating surface shared by the lysis liquid bin, the magnetic bead liquid bin 5, the washing liquid bin 6, the elution liquid bin 7 and the straw cavity 2 respectively, the needle assembly 8 is used to pierce the separating surface to make the corresponding reagent bin (lysis liquid bin, magnetic bead liquid bin 5, washing liquid bin 6, elution liquid bin 7) and the straw cavity 2 communicate with each other, so that the reagent in the reagent bin can enter the straw cavity 2, and a magnetic sheet slot 9 is further arranged on the outer wall surface of the straw cavity 2, and a magnetic sheet 10 can be inserted into the magnetic sheet slot 9. In this embodiment, the reagents required for lysis, combination, washing and elution steps of the nucleic acid sample, such as guanidine salt, magnetic bead liquid (magnetic bead + deionized water), washing liquid and elution liquid, are stored in the corresponding reagent bins in advance and integrated with the straw cavity 2, which is convenient for carrying to the outdoor for nucleic acid extraction operation; in the extraction process, the straw cavity 2 is pressed first, and the bottom end of the inlet and outlet pipe 1 is inserted into the nucleic acid sample liquid surface, after the hand is released, the nucleic acid sample is sucked into the straw cavity 2 along the inlet and outlet pipe 1, then the switch valve 3 is closed, the needle assembly 8 in the corresponding reagent bin is squeezed, the needle assembly 8 pierces the separating surface to make the corresponding reagent bin and the interior of the straw cavity 2 communicate with each other, and the reagent stored in the reagent bin is squeezed into the straw cavity 2, so that the corresponding reagent can be added to perform the corresponding extraction step, since the reagent does not need to be transferred by using a pipette, there is no risk of reagent dripping outside, and the operation is very simple and fast, in the extraction process, the magnetic sheet 10 is inserted into the magnetic sheet slot 9 or taken out to adsorb and release the magnetic beads, which effectively improves the convenience of extraction operation, speeds up the nucleic acid extraction process and improves the extraction efficiency.

[0023] In the preferred embodiment, the inlet and outlet tube 1 is provided with scale lines 11. The scale lines 11 are arranged along the length direction of the inlet and outlet tube 1, and the scale lines 11 facilitate the control of the amount of nucleic acid sample sucked in.

[0024] In the specific example, the switch valve 3 comprises a valve rod 301 and a ball 302. The ball 302 is arranged inside the inlet and outlet tube 1, and the valve rod 301 is rotatable to penetrate into the interior of the inlet and outlet tube 1 and is connected with the ball 302. The valve rod 301 is provided with a rotating handle 303 at one end outside the inlet and outlet tube 1. Specifically, the ball 302 is provided with a channel, and the channel is perpendicular to the valve rod 301. When the rotating handle 303 drives the valve rod 301 and the ball 302 to rotate to a certain angle, the channel can communicate with the inlet and outlet tube 1. Conversely, the ball 302 blocks and closes the inlet and outlet tube 1.

[0025] In the preferred embodiment, the top of the suction tube cavity 2 is provided with a gas vent 202, which is in communication with the interior of the suction tube cavity 2. The gas vent 202 is provided with a cap 203, which is used to block the gas vent 202 to avoid the direct communication between the suction tube cavity 2 and the external atmosphere. When it is necessary to discharge the liquid in the suction tube cavity 2, the switch valve 3 is first opened, and then the suction tube cavity 2 is pressed to make the liquid flow out from the bottom end of the inlet and outlet tube 1. When the amount of liquid in the suction tube cavity 2 is relatively large, there may be some liquid remaining in the inlet and outlet tube 1 after the suction tube cavity 2 is completely pressed. If the suction tube cavity 2 is released, the liquid may be sucked back, and multiple pressing operations are required, which is low in efficiency. By opening the cap 203 of the gas vent 202, the air pressure in the suction tube cavity 2 is balanced with the external atmosphere, and the liquid can automatically flow out under the action of gravity, effectively avoiding the liquid remaining in the suction tube cavity 2.

[0026] In the specific example, the piercing needle assembly 8 is fixed on the bottom surface of the lysis liquid tank 4, the magnetic bead liquid tank 5, the washing liquid tank 6 and the elution liquid tank 7 respectively, the piercing needle assembly 8 comprises a U-shaped elastic sheet 801 and a puncture needle 802, the U-shaped elastic sheet is a U-shaped structure composed of two sheet bodies, the lower wall of the U-shaped elastic sheet 801 is fixedly connected with the bottom surface of the corresponding lysis liquid tank 4, the magnetic bead liquid tank 5, the washing liquid tank 6 and the elution liquid tank 7 after being turned over by 90°, a puncture hole 803 is arranged on the lower wall of the U-shaped elastic sheet 801, the puncture needle 802 is arranged in the opening of the U-shaped elastic sheet 801 and is fixedly connected with the upper side wall of the U-shaped elastic sheet 801, and the puncture needle 802 corresponds to the puncture hole 803; when it is needed to puncture to make the corresponding reagent tank communicate with the straw cavity 2, the lower wall of the U-shaped elastic sheet 801 is moved downward by pressing the lower wall, the puncture needle 802 moves downward synchronously to pierce the separation surface at the puncture hole 803, and the puncture needle 802 is retracted from the puncture hole 803 after being released, so that the corresponding reagent tank can communicate with the straw cavity 2, by adopting the above structure, the puncture needle 802 can only pierce the separation surface in the region of the puncture hole 803 and cannot pierce other surfaces, so that the puncture needle 802 can avoid piercing other surfaces to cause leakage of the corresponding reagent tank, and the puncture needs to overcome the elastic force between the U-shaped elastic sheets 801, thereby reducing the risk of liquid leakage caused by accidental puncture of the nucleic acid extraction kit during transportation.

[0027] In the preferred embodiment, the nucleic acid extraction kit further comprises a PCR tube 12 corresponding to the bottom end of the inlet and outlet liquid pipe 1, the PCR tube 12 is used for collecting the extracted nucleic acid extraction liquid, and the nucleic acid extraction liquid is conveniently transferred to the instrument for detection.

[0028] When the nucleic acid extraction kit is used, the bottom end of the inlet and outlet tube 1 is inserted into the liquid surface of the nucleic acid sample after pressing the straw cavity 2, then the nucleic acid sample is sucked into the straw cavity 2 by hand, the switch valve 3 is closed, the needle assembly 8 in the lysis solution tank 4 is squeezed, the needle assembly 8 pierces the separation surface to make the lysis solution tank 4 and the inside of the straw cavity 2 communicate, then the high salt solution (guanidine salt) stored in the lysis solution tank 4 is squeezed into the straw cavity 2, and the nucleic acid DNA / RNA in the sample is lysed and separated; then the needle assembly 8 in the magnetic bead liquid tank 5 is squeezed to pierce the separation surface to make the magnetic bead liquid tank 5 and the inside of the straw cavity 2 communicate, and the magnetic bead liquid is squeezed into the straw cavity 2 for mixing, the nucleic acid DNA / RNA in the mixed solution combines with the nano magnetic beads in the magnetic bead liquid, under the action of the external magnetic field, the magnetic bead-nucleic acid complex is formed, the magnetic sheet 10 is inserted into the magnetic sheet slot 9, the magnetic bead-nucleic acid complex is concentrated and adsorbed on the side wall of the straw cavity 2 in the magnetic sheet slot 9 area, then the switch valve 3 is opened to discharge the mixed solution in the straw cavity 2, after the discharge is completed, the switch valve 3 is closed, the magnetic sheet 10 can be taken out, the needle assembly 8 in the washing liquid tank 6 is squeezed to pierce the separation surface, then the washing liquid is squeezed into the straw cavity 2 for washing, the impurities on the magnetic bead-nucleic acid complex are removed, after the washing step is completed, the magnetic sheet 10 is inserted again and the impurity liquid is discharged, after the impurity liquid is discharged, the switch valve 3 is closed, the needle assembly 8 in the eluent tank 7 is squeezed to pierce the separation surface, then the eluent is squeezed into the straw cavity 2 for the elution step, after the elution step is completed, the magnetic sheet 10 is inserted for magnetic separation, then the extracted and purified DNA solution is discharged into the PCR tube 12, and the nucleic acid extraction is completed.

[0029] The above is further detailed description of the utility model in combination with specific / preferred embodiments, and cannot be deemed as limitation of the specific implementation of the utility model to these descriptions. For ordinary skilled in the art to which the utility model belongs, without departing from the concept of the utility model, some substitutions or variations can be made to the described embodiments, and these substitutions or variations shall be deemed as falling within the protection scope of the utility model. In the description of the specification, the description of the reference terms "an embodiment", "some embodiments", "preferred embodiment", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in combination with the embodiment or example are contained in at least one embodiment or example of the utility model. In the specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In the case of not mutually contradictory, the skilled in the art can combine and combine the different embodiments or examples described in the specification and the features of different embodiments or examples. Although the embodiments of the utility model and its advantages have been described in detail, it should be understood that various changes, substitutions and modifications can be made in this paper without departing from the protection scope of the patent application.

Claims

1. A nucleic acid extraction kit characterized by: The application relates to a liquid inlet and outlet pipe and a straw cavity, wherein the straw cavity is made of rubber material, the straw cavity is communicated with the top end of the liquid inlet and outlet pipe, a switch valve is arranged on the liquid inlet and outlet pipe, a lysing liquid bin, a magnetic bead liquid bin, a washing liquid bin and an elution liquid bin are respectively arranged on the straw cavity, a needle assembly is arranged on the separation surface of the straw cavity, a magnetic sheet slot is arranged on the outer wall surface of the straw cavity, and a magnetic sheet can be inserted into the magnetic sheet slot.

2. The nucleic acid extraction kit according to claim 1, wherein: The liquid inlet and outlet pipe is provided with scale lines.

3. The nucleic acid extraction kit as described in claim 1, characterized in that: The liquid inlet and outlet pipe is made of plastic or glass.

4. The nucleic acid extraction kit according to claim 1, wherein: The inner wall surface of the straw cavity is provided with an anticorrosive layer.

5. The nucleic acid extraction kit as described in claim 4, characterized in that: The anticorrosive layer is made of a polytetrafluoroethylene coating.

6. The nucleic acid extraction kit according to claim 1, wherein: The switch valve comprises a valve rod and a ball, the ball is arranged in the liquid inlet and outlet pipe, the valve rod can be rotated to penetrate into the liquid inlet and outlet pipe and is connected with the ball, and a rotating handle is arranged on one end of the valve rod which is located outside the liquid inlet and outlet pipe.

7. The nucleic acid extraction kit according to claim 1, characterized in that: A gas vent is arranged on the top of the straw cavity, and a plug cap is arranged at the gas vent.

8. The nucleic acid extraction kit according to any one of claims 1 to 7, wherein: The needle assemblies are respectively fixedly arranged on the bottom surfaces of the lysing liquid bin, the magnetic bead liquid bin, the washing liquid bin and the elution liquid bin.

9. The nucleic acid extraction kit as described in claim 8, characterized in that: The needle assembly comprises a U-shaped elastic sheet and a puncture needle, the lower wall surface of the U-shaped elastic sheet is fixedly connected with the bottom surfaces of the lysing liquid bin, the magnetic bead liquid bin, the washing liquid bin and the elution liquid bin after being turned over by 90 degrees, a puncture hole is arranged on the lower wall surface of the U-shaped elastic sheet, the puncture needle is arranged in the opening of the U-shaped elastic sheet and is fixedly connected with the upper side wall of the U-shaped elastic sheet, and the puncture needle corresponds to the puncture hole.

10. The nucleic acid extraction kit as described in claim 9, characterized in that: The application further comprises a PCR tube which corresponds to the bottom end of the liquid inlet and outlet pipe.

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