A DNA extraction kit for bone samples

Abstract

This invention belongs to the field of reagent kit technology, specifically relating to a DNA extraction kit for bone samples. The kit includes a housing with a separation and stirring tank, a magnetic bead storage tank, and at least one liquid storage tank. A filter membrane is installed within the separation and stirring tank, dividing it into a first chamber and a second chamber, with the first chamber having a smaller space than the second chamber. This invention eliminates the need for centrifugation, directly separating bone powder and DNA for subsequent extraction steps. This not only avoids DNA loss caused by centrifugation but also shortens operation time, facilitates automation, simplifies the operation, and reduces the risk of sample contamination.

Description

Technical Field

[0001] This utility model relates to a DNA extraction kit for bone samples, belonging to the field of reagent kit technology. Background Technology

[0002] Bone (including teeth) is a special type of biological sample, but due to the unique structure of bone tissue and the influence of environmental factors, bone DNA extraction is relatively difficult. To facilitate the release of DNA from bone tissue, a decalcification step is generally added during the extraction process. EDTA is used for decalcification to remove calcium from hydroxyapatite (the main inorganic component of human and animal bones). Studies have shown that decalcification has a significant impact on bone DNA extraction. Decalcification facilitates cell dispersion and assists the lysis buffer in disrupting cell and nuclear membranes, pulling histones away from DNA molecules and increasing DNA yield. Currently, reagents for DNA extraction from bone samples require centrifugation after decalcification to extract DNA from the supernatant. During centrifugation, bone powder particles may adhere to a large amount of DNA, which settles with the bone powder particles, resulting in significant DNA loss after centrifugation and affecting the sample volume of the kit. Therefore, a bone sample DNA extraction kit that can avoid DNA loss caused by centrifugation is needed. Utility Model Content

[0003] This invention addresses the aforementioned technical problems in the prior art by providing a DNA extraction kit for bone samples.

[0004] The technical solution of this utility model to solve the above-mentioned technical problems is as follows: A DNA extraction kit for bone samples includes a box body, the box body is provided with a separation stirring tank, a magnetic bead storage tank and at least one liquid storage tank, the separation stirring tank is provided with a filter membrane, the filter membrane divides the separation stirring tank into a first tank chamber and a second tank chamber, the space of the first tank chamber is smaller than the space of the second tank chamber.

[0005] The beneficial effects of this invention are as follows: This invention can directly separate bone powder and DNA without centrifugation for subsequent extraction steps, which not only avoids DNA loss caused by centrifugation, but also shortens the operation time, makes it easier to automate, simplifies the operation steps, and reduces the risk of sample contamination.

[0006] Based on the above technical solution, the present invention can be further improved as follows:

[0007] Furthermore, the filter membrane is detachably installed inside the separation stirring tank.

[0008] The beneficial effect of adopting the above-mentioned further technical solution is that the filter membrane can be detachably installed in the separation stirring tank, which facilitates the use of the reagent kit.

[0009] Furthermore, the filter membrane is installed inside the separation and stirring tank via a membrane clamping frame.

[0010] Furthermore, there are two membrane frames, and the filter membrane is disposed between the two membrane frames.

[0011] The beneficial effects of adopting the above-mentioned further technical solutions are: the filter membrane is installed by the membrane clamping frame, which makes the installation and disassembly of the filter membrane in the separation and stirring tank very convenient. The two membrane clamping frames can be clamped together by heat sealing, and the filter membrane is held between the two membrane clamping frames, which is easy to process.

[0012] Furthermore, the inner wall of the separating stirring tank is provided with a slot, and the clamping frame is provided with an insertion edge corresponding to the slot, and the clamping frame is inserted into the slot.

[0013] The beneficial effects of adopting the above-mentioned further technical solutions are: the membrane frame is installed through a slot, which is simple to operate and easy to install and disassemble.

[0014] Furthermore, the pore size of the filter membrane is 50μm-150μm.

[0015] The beneficial effect of adopting the above-mentioned further technical solution is that the filter membrane with a pore size of 50μm-150μm can prevent most of the bone powder from entering the second chamber, thus avoiding the bone powder from affecting the extraction of DNA.

[0016] Furthermore, the clamping frame is provided with an inverted T-shaped pressing strip.

[0017] The beneficial effect of adopting the above-mentioned further technical solutions is that the membrane pressing strip presses the filter membrane more evenly, avoiding unevenness of the filter membrane that would affect the filtration effect.

[0018] Furthermore, the liquid storage tank is provided with at least four.

[0019] The beneficial effects of adopting the above-mentioned further technical solutions are: by setting up multiple liquid storage tanks for storing various liquids required for DNA extraction, such as washing solutions, or for storing waste liquids after the experiment, the use of the kit is facilitated and the structure of the kit is simplified. Attached Figure Description

[0020] Figure 1 This is a three-dimensional structural diagram of the present invention;

[0021] Figure 2 This is a schematic diagram of the process of inserting the membrane frame of this utility model into the separation and stirring tank;

[0022] Figure 3 This is a top view of the present invention;

[0023] Figure 4 This is a three-dimensional sectional view of the present invention;

[0024] Figure 5 This is a schematic diagram of the structure of the membrane frame and filter membrane of this utility model;

[0025] Figure 6 This is the front view of the present invention;

[0026] Figure 7 This is a cross-sectional view of the present invention.

[0027] The attached diagram is labeled as follows: 1. Separation stirring tank; 101. First tank chamber; 102. Second tank chamber; 103. Slot; 2. Magnetic bead storage tank; 3. Liquid storage tank; 4. Filter membrane; 5. Membrane clamping frame; 501. Membrane pressing strip; 6. Box body. Detailed Implementation

[0028] The principles and features of this utility model are described below with reference to the accompanying drawings. The examples given are only for explaining this utility model and are not intended to limit the scope of this utility model.

[0029] See Figures 1-7 A DNA extraction kit for bone samples includes a box body 6. The box body 6 is provided with a separation stirring tank 1, a magnetic bead storage tank 2 and at least one liquid storage tank 3. The separation stirring tank 1 is pre-filled with a binding solution, and the magnetic bead storage tank 2 is pre-filled with magnetic beads. The separation stirring tank 1 is provided with a filter membrane 4, which divides the separation stirring tank 1 into a first chamber 101 and a second chamber 102. The space of the first chamber 101 is smaller than the space of the second chamber 102. Specifically, the filter membrane 4 is vertically arranged in the separation stirring tank 1, dividing the separation stirring tank 1 into the first chamber 101 and the second chamber 102 along the length of the box body 6. The length of the first chamber 101 is 1 / 2 to 1 / 5 of the length of the second chamber 102.

[0030] The filter membrane 4 is detachably installed inside the separation and stirring tank 1. See details below. Figures 2-5 In this embodiment, the filter membrane 4 is installed in the separation and stirring tank 1 via a clamping frame 5. The inner wall of the separation and stirring tank 1 is provided with a slot 103, and the clamping frame 5 has an insertion edge corresponding to the slot 103. The clamping frame 5 is inserted into the slot 103. Two clamping frames 5 are provided, and the two clamping frames 5 are clamped and heat-sealed together, with the filter membrane 4 positioned between the two clamping frames 5. An inverted T-shaped pressure strip 501 is provided on the clamping frame 5, dividing the filter membrane 4 into three regions, making the filter membrane 4 flatter and improving its filtration effect. The pore size of the filter membrane 4 is 50μm-150μm; specifically, in this embodiment, the pore size of the filter membrane 4 is 100μm.

[0031] At least four liquid storage tanks 3 are provided. Specifically, in this embodiment, five liquid storage tanks 3 are provided, of which three liquid storage tanks 3 are pre-filled with washing solution, one liquid storage tank 3 is pre-filled with eluent, and one liquid storage tank 3 is used for pre-filling other reagents required for the experiment or storing waste liquid after the experiment. After the separation stirring tank 1, magnetic bead storage tank 2 and liquid storage tank 3 are pre-filled with reagents, they are heat-sealed with aluminum foil.

[0032] This invention divides the separation and stirring tank into a first chamber 101 and a second chamber 102 using a filter membrane. The separation and stirring tank 1 is pre-filled with a binding solution. When separating bone powder and DNA, the decalcified bone powder solution is added to the first chamber 101. The bone powder solution mixes with the binding solution through the filter membrane 4. The filter membrane 4 effectively isolates the bone powder from entering the second chamber 102. After adding magnetic beads to the second chamber 102, the magnetic rod is used to repeatedly oscillate and stir the bone powder in the second chamber 102. The mixture of decalcified solution and binding solution can repeatedly wash the bone powder, fully releasing the DNA and allowing the magnetic beads to fully adsorb the DNA. At the same time, the filter membrane 4 also isolates most of the bone powder particles from entering the second chamber 102.

[0033] The steps for using this utility model are as follows:

[0034] 1. Weigh 1g of bone meal and add 0.5M EDTA for decalcification;

[0035] 2. Add the decalcified bone meal solution into the first chamber 101 of the separation and stirring tank 1;

[0036] 3. Place the box 6 in a semi-automatic or fully automatic device, and the machine will adsorb the magnetic beads in the magnetic bead storage tank 2 into the second chamber 102 of the separation stirring tank 1.

[0037] 4. Stir thoroughly with the magnetic rod and incubate at room temperature for 15-20 minutes;

[0038] 5. Sequentially adsorb the magnetic beads into the three liquid storage tanks 3, and wash the magnetic beads three times with the washing liquid in the liquid storage tanks 3;

[0039] 6. Adsorb the magnetic beads into the fourth liquid storage tank 3, and elute the DNA adsorbed on the magnetic beads with the elution solution in the liquid storage tank 3. Mix with the magnetic rod and elute at 65℃-80℃.

[0040] 7. After magnetic attraction, discard the magnetic beads and collect the DNA from the elution solution in the fourth liquid storage tank 3.

[0041] The above description is only a preferred embodiment of the present utility model and is not intended to limit the present utility model. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present utility model should be included within the protection scope of the present utility model.

Claims

1. A DNA extraction kit for bone samples, comprising a housing (6), characterized in that, The box body (6) is provided with a separation stirring tank (1), a magnetic bead storage tank (2) and at least one liquid storage tank (3). The separation stirring tank (1) is provided with a filter membrane (4). The filter membrane (4) divides the separation stirring tank (1) into a first tank chamber (101) and a second tank chamber (102). The space of the first tank chamber (101) is smaller than the space of the second tank chamber (102).

2. The DNA extraction kit for skeletal samples according to claim 1, characterized in that, The filter membrane (4) is detachably installed in the separation stirring tank (1).

3. The DNA extraction kit for bone samples according to claim 2, characterized in that, The filter membrane (4) is installed in the separation stirring tank (1) through the membrane clamping frame (5).

4. The DNA extraction kit for bone samples according to claim 3, characterized in that, Two membrane frames (5) are provided, and the filter membrane (4) is located between the two membrane frames (5).

5. The DNA extraction kit for bone samples according to claim 4, characterized in that, The inner wall of the separation stirring tank (1) is provided with a slot (103), and the clamping frame (5) is provided with an insertion edge corresponding to the slot (103), and the clamping frame (5) is inserted into the slot (103).

6. The DNA extraction kit for bone samples according to claim 5, characterized in that, The pore size of the filter membrane (4) is 50μm-150μm.

7. The DNA extraction kit for skeletal samples according to claim 6, characterized in that, The clamping frame (5) is provided with an inverted T-shaped pressing strip (501).

8. The DNA extraction kit for bone samples according to claim 7, characterized in that, The liquid storage tank (3) is provided with at least four.

Images