Miniature nucleic acid extraction device supporting temperature adjustment and extraction pen

Abstract

The invention relates to the technical field of nucleic acid extraction, in particular to a micro nucleic acid extraction device supporting temperature adjustment and an extraction pen, the micro nucleic acid extraction device comprises a nucleic acid extraction heating sleeve, a temperature adjustment cavity is formed in the nucleic acid extraction heating sleeve, the nucleic acid extraction heating sleeve comprises a sleeve body, and reagent tube groove holes are formed in the two side edge ends of the sleeve body. The test tube is correspondingly heated by matching with the heating groove holes in the groove holes and the electric heating pieces, a five-gear heating function is achieved, cells can be better split, nucleic acid is released and DNA elution is accelerated through heating, the test tube is matched with an extraction pen for use, magnetic beads carrying nucleic acid are adsorbed by a magnetic rod and moved into reagent tubes with different purposes, the effects of uniform mixing and stirring are achieved, and the test tube is convenient to use. The nucleic acid sample pretreatment device can improve the convenience of operations such as transfer, washing and elution of magnetic beads in the nucleic acid extraction process, is simple in structure, simplifies experimental steps, can be carried and used with other nucleic acid detection products, and is easier to use in various severe environments.

Description

Technical Field

[0001] The present invention relates to the technical field of nucleic acid extraction, in particular to a micro nucleic acid extraction device and an extraction pen supporting temperature regulation. Background Art

[0002] In the field of genetic engineering and protein engineering research, nucleic acid molecules are the primary research subjects, and the separation and linearization of nucleic acids are the basic techniques for nucleic acid research. Nucleic acids are the basic units that represent the genetic characteristics of living organisms. Nucleic acid testing is an advanced biological test performed at the molecular level. Compared with traditional morphological testing, cytological testing, immunological testing, etc., it has significant advantages such as greater sensitivity, specificity, and no window period. Nucleic acid testing includes qualitative PCR, molecular hybridization, real-time fluorescence quantitative PCR and other technologies. The primary key to these nucleic acid testing technologies is to complete the extraction of nucleic acids from biological samples. Therefore, the effective and accurate extraction of nucleic acid templates has become a prerequisite for subsequent nucleic acid testing. Domestic nucleic acid extraction mainly uses traditional manual preparation methods combined with instrumentation for extraction operations.

[0003] In the process of nucleic acid extraction, the magnetic bead extraction method is mainly used in conjunction with instruments for extraction. The magnetic rod method uses the transfer of magnetic beads and the fixation of liquid to achieve the separation of nucleic acids. It has the same principle and process as the aspiration method, but the way of separating magnetic beads and liquid is different. The magnetic rod method separates the magnetic beads from the waste liquid by adsorbing them on the magnetic rod, and then puts them into the liquid in the next step to achieve the extraction of nucleic acids. Functionalized magnetic beads are used to capture nucleic acids through interaction with nucleic acids, and are quickly extracted by magnetic separation. The main steps include lysing the sample, adding magnetic beads for incubation to adsorb nucleic acids, magnetic separation of adsorbed magnetic beads, washing to remove impurities, and eluting to obtain nucleic acids.

[0004] The above-mentioned extraction method needs to be used in conjunction with a nucleic acid extraction instrument. Its advantages are rapid extraction, specificity, and automation, but its disadvantages are the need for dedicated instruments and consumables. At the same time, the instrument is large and heavy, making it difficult to carry around. In addition, the pretreatment of nucleic acid samples is time-sensitive, and the nucleic acid transfer process of magnetic beads is cumbersome and complex, making it extremely inconvenient in harsh working environments.

[0005] In view of the above problems, we propose a micro nucleic acid extraction device and an extraction pen that support temperature regulation. Summary of the Invention

[0006] The object of the present invention is to provide a micro nucleic acid extraction device and an extraction pen that support temperature regulation, so as to solve the problems raised in the above background technology.

[0007] To achieve the above-mentioned purpose, the present invention provides the following technical solution: an extraction pen, comprising a nucleic acid extraction pen, the nucleic acid extraction pen comprising a pen shaft, and the bottom end of the pen shaft is provided with a magnetic rod tube sleeve, a magnetic head is provided inside the magnetic rod tube sleeve, and the top of the magnetic head is connected to a magnetic rod, a stop component is provided inside the top end of the pen shaft, and a spring is attached to the bottom end of the stop component, and the top end of the stop component is connected to a push lever.

[0008] Furthermore, the magnetic rod is connected to the magnetic head and the stopping component, and the stopping component forms an elastic telescopic structure with the pen body through a spring.

[0009] Furthermore, a micro nucleic acid extraction device supporting temperature control includes a nucleic acid extraction heating jacket, a temperature control chamber is provided inside the nucleic acid extraction heating jacket, the nucleic acid extraction heating jacket includes a jacket body, a power interface is provided at the bottom of one end of the jacket body, a power cord is plugged into one side of the power interface, a temperature control button is provided at the top of the power interface, a first slot is provided at both side ends of the jacket body, a second slot is provided longitudinally of the first slot, a third slot is provided longitudinally of the second slot, a fourth slot is provided longitudinally of the third slot, a fifth slot is provided longitudinally of the fourth slot, and a sixth slot is provided longitudinally of the fifth slot;

[0010] Reagent tubes are provided in the first to sixth slots described above, and the reagent tubes are plastic outer spiral capped cone-bottomed freezing tubes with a tube volume of 1.5-2 mL.

[0011] Furthermore, the first to sixth slots are symmetrically arranged along both side ends of the sleeve, and the first to sixth slots are equidistantly arranged along the side ends of the sleeve.

[0012] Furthermore, the temperature adjustment buttons are symmetrically distributed along the middle of both ends of the sleeve, and the temperature adjustment buttons and the power interface are connected to the temperature adjustment cavity.

[0013] Furthermore, the temperature control chamber includes a cavity, and a battery compartment is provided at the middle end of the cavity, two ends of the battery compartment are connected to a thermostat, and one side of the thermostat at one end is connected to a temperature control button, the other side of the thermostat is connected to a processor, and one side of the processor is connected to a Bluetooth module, and electric heaters are distributed on both sides of the cavity, and the electric heaters are engaged with the inner walls of the first slot and the fifth slot;

[0014] The above-mentioned electric heater is an aluminum metal heat conductive plate. The temperature adjustment button and thermostat are respectively arranged on both sides of the nucleic acid extraction heating jacket, and are provided with different temperature levels, from level four to level eight, for the heating temperature during the nucleic acid lysis and elution steps.

[0015] Furthermore, the electric heater is connected to the battery compartment and the thermostat wire, and the thermostat is symmetrically distributed along both ends of the cavity.

[0016] Furthermore, the processor is connected to the thermostat and the Bluetooth module via wires.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] In the present invention, the slots on both sides of the nucleic acid extraction heating jacket are grouped into groups of six, which is convenient for comparison and use. The heating slots and electric heaters in the slots are used to heat the test tubes accordingly. The jacket has five heating levels, which can better lyse cells, release nucleic acids, and accelerate DNA elution through heating. The overall structure is lightweight. As a nucleic acid sample pretreatment device, it not only has a simple structure and simplifies the experimental steps, but can also be carried and used with other nucleic acid detection products, making it easier to use in a variety of harsh environments.

[0019] In the present invention, in order to facilitate the use of the heating process and temperature control, when the device is used for nucleic acid extraction and heating, the temperature control button on both sides can be used to trigger the thermostat to control the temperature of the electric heater. After the battery compartment is connected to the power cord, the connected electric heater is powered and heated. During the temperature control process, different temperature levels can be set to adjust the heating temperature during the nucleic acid lysis and elution steps. At the same time, the built-in Bluetooth module is connected to the mobile phone terminal, and the heating can be controlled by the mobile phone terminal or the button. The mobile phone connects to the device via Bluetooth, sets the corresponding parameters, and then performs a stable heating extraction operation.

[0020] In the present invention, in order to improve the convenience of operations such as transferring, washing and eluting magnetic beads during nucleic acid extraction and accelerate the nucleic acid extraction process, the device is used in conjunction with the nucleic acid extraction pen. The stop component can be activated by pressing the push lever, and the magnetic rod connected to the stop component can be kept in an extended state by pressing down and retracting. The magnetic rod is used to absorb the magnetic beads carrying nucleic acid and move them to reagent tubes for different purposes, thereby achieving the effect of mixing and stirring. The convenience of operations such as transferring, washing and eluting magnetic beads during nucleic acid extraction can be improved, and the nucleic acid extraction process can be accelerated. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a schematic diagram of the three-dimensional structure of the nucleic acid extraction heating jacket of the present invention;

[0022] Figure 2 This is a schematic diagram of the top view of the nucleic acid extraction heating jacket of the present invention;

[0023] Figure 3 This is a schematic diagram of the top view of the temperature adjustment chamber in the nucleic acid extraction heating jacket of the present invention;

[0024] Figure 4 This is a schematic side view of the nucleic acid extraction heating jacket of the present invention;

[0025] Figure 5 This is a schematic diagram of the three-dimensional structure of the nucleic acid extraction pen of the present invention;

[0026] Figure 6 This is a schematic diagram of the three-dimensional structure of the magnetic rod in the nucleic acid extraction pen of the present invention;

[0027] Figure 7 This is a schematic diagram of the internal structure of the nucleic acid extraction pen of the present invention.

[0028] In the picture:

[0029] 1. Nucleic acid extraction heating jacket; 101. Jacket body; 102. Power interface; 103. Power cord; 104. Temperature adjustment button; 105. First slot; 106. Second slot; 107. Third slot; 108. Fourth slot; 109. Fifth slot; 110. Sixth slot.

[0030] 2. Thermostatic chamber; 201. Cavity; 202. Battery compartment; 203. Thermostat; 204. Processor; 205. Bluetooth module; 206. Heating element.

[0031] 3. Nucleic acid extraction pen; 301. Pen body; 302. Magnetic rod sleeve; 303. Magnetic head; 304. Magnetic rod; 305. Stop component; 306. Spring; 307. Push lever. DETAILED DESCRIPTION

[0032] To facilitate solving the problem, the embodiments of the present invention provide a micro-nucleic acid extraction device and extraction pen that support temperature regulation. The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] like Figure 1-2 As shown, a micro nucleic acid extraction device supporting temperature control also includes a nucleic acid extraction heating jacket 1, the nucleic acid extraction heating jacket 1 is provided with a temperature control chamber 2, the nucleic acid extraction heating jacket 1 includes a jacket body 101, a power interface 102 is provided at the bottom of one end of the jacket body 101, and a power cord 103 is plugged into one side of the power interface 102, and a temperature control button 104 is provided on the top of the power interface 102. First slots 105 are provided on both sides of the jacket body 101, and second slots 106 are longitudinally provided in the first slot 105, and third slots 107 are longitudinally provided in the second slot 106, and fourth slots 108 are longitudinally provided in the third slot 107, and fifth slots 109 are longitudinally provided in the fourth slot 108, and sixth slots 110 are longitudinally provided in the fifth slot 109.

[0034] In order to provide a small, light and temperature-adjustable micro nucleic acid extraction device to solve the timeliness and complexity problems of nucleic acid sample pre-treatment, the magnetic beads can be easily washed and eluted during the nucleic acid transfer process, and the operation is simple, fast and efficient. Figure 1-2 As shown, the housing 101 of the nucleic acid extraction heating jacket 1 is made of plastic and can be shaped like a cuboid, a cube, a cylinder, etc., with cylindrical slots symmetrically arranged at both ends, which can be used to adjust the temperature in conjunction with two temperature adjustment buttons 104 on the side, and the power interface 102 is connected to the power cord 103 to provide power support for the internal battery compartment 202, and the six reagent tubes for different purposes built into the slots, including lysis, binding, wash one, wash two, elution, and sample bottle, correspond to the first slot 105, the second slot 106, and the second slot 106 respectively. , the third slot 107, the fourth slot 108, the fifth slot 109 and the sixth slot 110. The slots on both sides of this device are grouped into six, which is convenient for comparison and use. The heating slots and the electric heating plate 206 in the slots are used to heat the test tubes accordingly. It has five heating functions, which can better lyse cells, release nucleic acids, and accelerate the elution of DNA through heating. The overall structure is light in weight. As a nucleic acid sample pretreatment device, it not only has a simple structure, but also simplifies the experimental steps. It can also be carried and used with other nucleic acid detection products, and is easier to use in a variety of harsh environments.

[0035] like Figure 3-4 As shown, the temperature control chamber 2 includes a cavity 201, and a battery compartment 202 is provided at the middle end of the interior of the cavity 201, and thermostats 203 are connected to both ends of the battery compartment 202, and one side of the thermostat 203 at one end is connected to the temperature control button 104, the other side of the thermostat 203 is connected to the processor 204, and one side of the processor 204 is connected to the Bluetooth module 205, and electric heating plates 206 are distributed on both sides of the cavity 201, and the electric heating plates 206 are engaged with the inner walls of the first slot 105 and the fifth slot 109. The electric heating plates 206 are aluminum metal thermal conductive plates, and the temperature control button 104 and the thermostat 203 are respectively arranged on both sides of the nucleic acid extraction heating jacket 1.

[0036] In order to facilitate the use of heating process and temperature control, such as Figure 3-4 As shown, when the device is used for heating during nucleic acid extraction, the temperature adjustment buttons 104 on both sides can be used to trigger the thermostat 203 to control the temperature of the electric heater 206. After the battery compartment 202 is connected to the power cord 103, the connected electric heater 206 is powered and heated. During the temperature adjustment process, different temperature levels can be set, which can be four to eight levels, for the heating temperature during nucleic acid lysis and elution steps. At the same time, the built-in Bluetooth module 205 is connected to the mobile phone terminal, and the heating can be controlled by the mobile phone terminal or the button. The mobile phone connects to the device via Bluetooth, sets the corresponding parameters, and then performs stable heating extraction operations.

[0037] like Figure 5-7 As shown, the present invention also provides an extraction pen, including a nucleic acid extraction pen 3, the nucleic acid extraction pen 3 includes a pen holder 301, and the bottom end of the pen holder 301 is provided with a magnetic rod sleeve 302, a magnetic head 303 is provided inside the magnetic rod sleeve 302, and the top of the magnetic head 303 is connected to a magnetic rod 304, a stopper 305 is provided inside the top end of the pen holder 301, and the bottom end of the stopper 305 is attached to a spring 306, and the top end of the stopper 305 is connected to a push lever 307;

[0038] In order to improve the convenience of transferring, washing and eluting magnetic beads during nucleic acid extraction and speed up the nucleic acid extraction process, such as Figure 5-7 As shown, the device is used in conjunction with the nucleic acid extraction pen 3. The stop component 305 can be activated by pressing the push lever 307, and the magnetic rod 304 connected to the stop component 305 is kept in an extended state by pressing down and retracting. During use, the magnetic beads are adsorbed on the magnetic suction head 303 extended from the magnetic rod tube sleeve 302. The magnetic rod 304 is used to adsorb the magnetic beads carrying nucleic acid and move them to reagent tubes for different purposes, thereby achieving the effect of mixing and stirring. This can improve the convenience of operations such as transferring, washing and eluting the magnetic beads during the nucleic acid extraction process, thereby accelerating the nucleic acid extraction process.

[0039] Finally, during use, the nucleic acid extraction heating sleeve 1 is connected to the power supply through the power cord 103, and the wheel light of the side temperature adjustment button 104 flashes, completing the startup. It is used in conjunction with the matching nucleic acid extraction kit and placed in the six slots in the specified order. The heating can be controlled by two methods: the mobile phone or the button. The mobile phone connects to the device through the Bluetooth module 205 and sets the corresponding parameters. For the button method, long press the temperature adjustment button 104 to wake up the heating gear, and then short press to switch between five temperature gears. The temperature adjustment buttons 104 on both sides respectively adjust the temperature of the lysis solution slot hole and the elution solution slot hole. The five temperature gears are adjustable, and the temperature parameters of each gear are shown in the following table. When the nucleic acid extraction pen 3 adsorbs the magnetic beads, the magnetic rod 304 is kept extended. When transferring to the next reagent tube, press the push lever 307 to put it in.

[0040] Temperature parameter table for each gear

[0041]

[0042] In summary, when the micro nucleic acid extraction device and the extraction pen are used, the operation process for nucleic acid extraction includes:

[0043] S1. Cracking: This step can be performed in the first slot 105:

[0044] (1) Add 200 μL of sample to the tube;

[0045] (2) Add 230 μL of Lys is Buffer I and freeze-dried beads of proteinase K and magnetic beads to the above tube, mix well, adjust the temperature to 70°C using the temperature control button 104, and incubate for 10 minutes;

[0046] (3) Use nucleic acid extraction pen 3 to adsorb the magnetic beads and transfer them to the next test tube;

[0047] S2. Combining: This step can be performed in the second slot 106:

[0048] (1) Add 320 μL Lysine Buffer II, mix well, and let stand at room temperature for 5 min (mix every 1 min);

[0049] (2) Use the nucleic acid extraction pen 3 to adsorb the magnetic beads and transfer them to the next test tube;

[0050] S3. Washing:

[0051] Washing buffer 1: This step can be performed in the third slot 107. Add 600 μL Washing Buffer I, shake and mix, and use the nucleic acid extraction pen 3 to adsorb the magnetic beads and then transfer;

[0052] Washing buffer II: This step can be performed in the fourth slot 108. Add 600uL Washing Buffer II, shake and mix, use the nucleic acid extraction pen 3 to adsorb the magnetic beads and transfer them to the next test tube;

[0053] Note: The washing solution should be removed as much as possible in the last washing step;

[0054] S4. Elution: This step can be performed in the fifth slot 109:

[0055] (1) After the magnetic beads are dried, add 50-100 μL of Elution Buffer and mix well, or use a pipette to slowly blow the magnetic beads to fully resuspend them, then adjust the temperature to 85°C using the temperature adjustment button 104 and incubate for 4 minutes;

[0056] (2) Use the nucleic acid extraction pen 3 to adsorb the magnetic beads and stir until the solution is clear. Transfer the supernatant to the sample bottle in the sixth slot 110. This is the purified genomic DNA, which can be used for subsequent experiments or stored at -20°C.

[0057] The lysis buffer used in the above operation process can be guanidine isothiocyanate, guanidine hydrochloride, sodium citrate, etc., the washing buffer can be 1M Tris-HCl, anhydrous ethanol, sodium chloride and EDTA, etc., and the eluent can be Tris, DEPC-treated water, etc.

[0058] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. An extraction pen, characterized in that: The invention comprises a nucleic acid extraction pen (3), wherein the nucleic acid extraction pen (3) comprises a pen holder (301), and the bottom end of the pen holder (301) is sleeved with a magnetic rod tube sleeve (302), a magnetic head (303) is arranged inside the magnetic rod tube sleeve (302), and the top of the magnetic head (303) is connected to a magnetic rod (304), a stop component (305) is arranged inside the top end of the pen holder (301), and the bottom end of the stop component (305) is affixed with a spring (306), and the top end of the stop component (305) is connected to a push lever (307).

2. The extraction pen according to claim 1, characterized in that: The magnetic rod (304) is connected to the magnetic head (303) and the stop component (305), and the stop component (305) forms an elastic telescopic structure with the pen holder (301) via a spring (306).

3. A micro nucleic acid extraction device supporting temperature regulation, comprising an extraction pen according to any one of claims 1 to 2, characterized in that: The invention comprises a nucleic acid extraction heating sleeve (1), wherein a temperature adjustment chamber (2) is provided inside the nucleic acid extraction heating sleeve (1), and the nucleic acid extraction heating sleeve (1) comprises a sleeve body (101), a power interface (102) is provided at the bottom of one end of the sleeve body (101), and a power cord (103) is plugged into one side of the power interface (102), and a temperature adjustment button (104) is provided at the top of the power interface (102), and a first slot hole (105) is provided at both side ends of the sleeve body (101), and the first slot hole (105) is longitudinally provided with a second slot hole (106), the second slot hole (106) is longitudinally provided with a third slot hole (107), and the third slot hole (107) is longitudinally provided with a fourth slot hole (108), the fourth slot hole (108) is longitudinally provided with a fifth slot hole (109), and the fifth slot hole (109) is longitudinally provided with a sixth slot hole (110); Reagent tubes are provided in the first slot (105) to the sixth slot (110) described above, and the reagent tubes are plastic outer spiral capped cone bottom freezing tubes, and the volume of the tubes is 1.5-2 mL.

4. The micro-nucleic acid extraction device supporting temperature regulation according to claim 3, characterized in that: The first slot hole (105) to the sixth slot hole (110) are symmetrically opened along the two side ends of the sleeve body (101), and the first slot hole (105) to the sixth slot hole (110) are equidistantly arranged along the side ends of the sleeve body (101).

5. The micro-nucleic acid extraction device supporting temperature regulation according to claim 3, characterized in that: The temperature adjustment buttons (104) are symmetrically distributed along the middle of both ends of the casing (101), and the temperature adjustment buttons (104) and the power interface (102) are connected to the temperature adjustment chamber (2).

6. The micro-nucleic acid extraction device supporting temperature regulation according to claim 3, characterized in that: The temperature adjustment chamber (2) comprises a chamber (201), and a battery compartment (202) is provided at the middle end of the chamber (201), two ends of the battery compartment (202) are connected to a thermostat (203), and one side of the thermostat (203) at one end is connected to a temperature adjustment button (104), the other side of the thermostat (203) is connected to a processor (204), and one side of the processor (204) is connected to a Bluetooth module (205), and electric heating plates (206) are distributed on both sides of the chamber (201), and the electric heating plates (206) are engaged with the inner walls of the first slot (105) and the fifth slot (109); The above-mentioned electric heating plate (206) is an aluminum metal heat conductive plate. The temperature adjustment button (104) and the thermostat (203) are respectively arranged on both sides of the nucleic acid extraction heating jacket (1), and are provided with different temperature levels, from level four to level eight, for the heating temperature during the nucleic acid lysis and elution steps.

7. The micro-nucleic acid extraction device supporting temperature regulation according to claim 6, characterized in that: The electric heater (206) is connected to the battery compartment (202) and the thermostat (203) via wires, and the thermostat (203) is symmetrically distributed along both ends of the cavity (201).

8. The micro-nucleic acid extraction device supporting temperature regulation according to claim 2, characterized in that: The processor (204) is connected to the thermostat (203) and the Bluetooth module (205) via wires.

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