Blood genome DNA extraction kit and extraction method

Abstract

The invention discloses a blood genome DNA extraction kit and an extraction method, and belongs to the technical field of nucleic acid extraction. The DNA extraction kit comprises a lysis solution, a first rinsing solution, a second rinsing solution and an eluent, wherein the lysate is prepared from the following components: 5.5 to 6 M of guanidine hydrochloride, 0.2 to 0.3 M of NaCl, 0.01 to 0.015 M of EDTA (Ethylene Diamine Tetraacetic Acid), 0.01 to 0.015 M of glucan, Triton X-100 with the volume concentration of 1.5 to 2 percent, 0.06 to 0.1 M of dithiothreitol, Tween-20 with the volume concentration of 5 to 10 percent and 10 to 50 mM of trehalose. The components of the lysate in the kit are optimized, through the synergistic effect of the components, the cell lysis efficiency is remarkably improved, meanwhile, DNA molecules higher in purity, more complete in structure and higher in extraction yield can be obtained, and therefore the kit has good application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of nucleic acid extraction, and in particular relates to a blood genome DNA extraction kit and an extraction method. Background Art

[0002] The quality of eukaryotic cell high molecular weight DNA extraction is an important factor affecting subsequent experiments, especially in the genotyping test for disease diagnosis. The high quality of template DNA is an important guarantee for the accuracy of the subsequent experimental results. In addition, the speed of extraction and whether the operation is simple are also the main factors considered by most laboratories. The high-purity and complete whole blood genomic DNA can be used for scientific research, clinical, and PCR laboratory rapid analysis requirements. Human DNA is of great significance for disease diagnosis or auxiliary diagnosis, genetic disease gene sequence analysis, biodiversity research, etc.

[0003] With the rapid development of modern molecular biology technology, kits for extracting high-purity blood DNA have been commercialized, but they are expensive and difficult to popularize in laboratories. At present, the methods commonly used in laboratories around the world to extract blood genomic DNA include the CTAB method, traditional DNA extraction methods, phenol-chloroform method and kit method. These methods have been proven to be feasible through a large number of experiments, but not all methods can extract blood genomic DNA with high purity. The main reason is that although there are a large number of red blood cells in mammalian blood, they do not have cell nuclei and can only be extracted from white blood cells with very little content in the blood. How to remove red blood cells from the blood without destroying white blood cells is the key to the success of the experiment.

[0004] At present, although kits for extracting genomic DNA from blood are widely used, existing kits for extracting genomic DNA from blood have problems such as low extraction efficiency and low purity of the extracted DNA. Summary of the invention

[0005] The purpose of the present invention is to provide a blood genomic DNA extraction kit and extraction method, which are used to solve the problems of low extraction efficiency and low purity of extracted DNA in existing blood genomic DNA extraction kits.

[0006] In a first aspect, the present invention provides a blood genomic DNA extraction kit, comprising a lysis solution, a first rinsing solution, a second rinsing solution and an elution solution; wherein the lysis solution comprises the following components: 5.5-6M guanidine hydrochloride, 0.2-0.3M NaCl, 0.01-0.015M EDTA, 0.01-0.015M dextran, Triton X-100 with a volume concentration of 1.5-2% (v / v), 0.06-0.1M dithiothreitol, Tween-20 with a volume concentration of 5-10% (v / v), and 10-50mM trehalose.

[0007] In the present invention, the inventors have found that guanidine hydrochloride in the lysis solution can destroy the cell membrane, denature and precipitate proteins, so that nucleic acids can get rid of the entanglement of proteins and be fully released, which is beneficial to the subsequent extraction process; at an appropriate concentration, NaCl can reduce the solubility of DNA in water, which is helpful for the precipitation and separation of DNA. While reducing the solubility of DNA, NaCl will not destroy the structure of DNA, thus ensuring the integrity of the extracted DNA; the main role of EDTA in genome extraction is to inhibit the activity of DNase (DNA enzyme) and prevent DNA degradation; dextran has good water solubility and biocompatibility, and can improve the purity of the extracted DNA; Triton X-100 can further destroy the cell membrane and release nucleic acid molecules; dithiothreitol can reduce and destroy disulfide bonds and reduce DNA dimerization, thereby improving the efficiency and purity of DNA extraction and providing high-quality DNA samples for subsequent molecular biology research; Tween-20 can combine with guanidine hydrochloride to effectively release DNA; trehalose can protect the integrity of the structure of the extracted nucleic acid molecules; the synergistic effect between the above components can not only significantly improve the efficiency of cell lysis, but also obtain DNA molecules with higher purity, more complete structure and higher extraction yield.

[0008] In the blood genomic DNA extraction kit provided by the present invention, the guanidine hydrochloride in the lysate can be, for example, 5.5M, 5.6M, 5.7M, 5.8M, 5.9M, 6M or other values ​​within the range; NaCl can be, for example, 0.2M, 0.22M, 0.25M, 0.28M, 0.3M or other values ​​within the range; EDTA can be, for example, 0.01M, 0.011M, 0.012M, 0.013M, 0.014M, 0.015M or other values ​​within the range; dextran can be, for example, 0.010M, 0.011M, 0.012M, 0.013M, 0.014M, 0.015M or other values ​​within the range; Triton The volume concentration (v / v) of X-100 can be, for example, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2% or other values ​​within this range; the volume concentration (v / v) of dithiothreitol can be, for example, 0.06M, 0.07M, 0.08M, 0.09M, 0.10M or other values ​​within this range; the volume concentration (v / v) of Tween-20 can be, for example, 5%, 6%, 7%, 8%, 9%, 10% or other values ​​within this range; the volume concentration of trehalose can be, for example, 10mM, 20mM, 30mM, 40mM, 50mM or other values ​​within this range.

[0009] In some embodiments, the first rinse solution includes the following components: 4.5-5M (for example, 4.5M, 4.6M, 4.7M, 4.8M, 4.9M, 5M or other values ​​within the range) guanidine hydrochloride, 10-15mM (for example, 10mM, 11mM, 12mM, 13mM, 14mM, 15mM or other values ​​within the range) Tris-HCl, 1-2mM (for example, 1mM, 1.2mM, 1.5mM, 1.8mM, 2mM or other values ​​within the range) EDTA, 0.1-0.3mM (for example, 0.1mM, 0.15mM, 0.2mM, 0.25mM, 0.3mM or other values ​​within the range) phenylmethylsulfonyl fluoride, PEG with a volume concentration of 10-25% (for example, 10%, 15%, 20%, 25% or other values ​​within the range, v / v) 8000.

[0010] In the present invention, the inventors further studied and found that guanidine hydrochloride in the first rinsing solution can further release DNA molecules; EDTA can further prevent DNA degradation; phenylmethylsulfonyl fluoride, as a protease inhibitor, can prevent DNA molecules from being degraded during the rinsing process, improve the extraction efficiency of DNA molecules and maintain their structural integrity; PEG 8000 can remove impurities and improve the purity of DNA molecules; the synergistic effect between the above substances can significantly improve the DNA elution efficiency and remove protein molecules in DNA.

[0011] In some embodiments, the second rinse solution includes the following components: 0.15-0.25M (e.g., 0.15M, 0.17M, 0.2M, 0.23M, 0.25M or other values ​​within the range) Tris, 0.01-0.02M (e.g., 0.01M, 0.012M, 0.015M, 0.018M, 0.02M or other values ​​within the range) SDS, 0.3-0.5M (e.g., 0.3M, 0.35M , 0.4M, 0.45M, 0.5M or other values ​​within this range) NaCl, 0.005-0.06M (for example, 0.005M, 0.01M, 0.03M, 0.05M, 0.06M or other values ​​within this range) EDTA, and sodium azide having a volume concentration of 0.01-0.03% (for example, 0.01%, 0.015%, 0.02%, 0.025%, 0.03% or other values ​​within this range, v / v).

[0012] In the present invention, the inventors further studied and found that SDS in the second rinse solution can combine with protein to form a precipitate, which helps to remove protein impurities in the sample and improve the purity of DNA; Tris can protect DNA from the influence of acidic substances or other degradation factors through its buffering effect, ensuring the quality of extracted DNA; by controlling the concentration of NaCl, DNA can be selectively precipitated, and at the same time, impurities such as proteins and polysaccharides in the solution can be removed, thereby improving the purity of DNA; sodium azide can further purify DNA; the above substances can significantly improve the elution efficiency through synergistic effects, and further improve the purity and concentration of DNA.

[0013] In some embodiments, the eluent includes the following components: 3-10 mM (for example, 3 mM, 5 mM, 7 mM, 9 mM, 10 mM or other values ​​within this range) Tris-HCl, 1-2 mM (for example, 1 mM, 1.2 mM, 1.5 mM, 1.8 mM, 2 mM or other values ​​within this range) EDTA, pH 7.2-8.2, for example, 7.2, 7.4, 7.6, 7.8, 8.2 or other values ​​within this range.

[0014] In some embodiments, the blood genomic DNA extraction kit also includes a proteinase K solution and a magnetic bead solution; wherein the concentration of the proteinase K solution is 80-150 μg / mL, for example, it can be 80 μg / mL, 100 μg / mL, 120 μg / mL, 150 μg / mL or other values ​​within this range; the concentration of the magnetic bead solution is 2-8 mg / mL, for example, it can be 2 mg / mL, 4 mg / mL, 6 mg / mL, 8 mg / mL or other values ​​within this range.

[0015] In a second aspect, the present invention provides a method for extracting blood genomic DNA using any of the above-mentioned extraction kits, comprising the following steps: S1, adding a lysis solution and a proteinase K solution to a blood sample to be extracted, oscillating and mixing, and then incubating to obtain a first mixed solution; S2, adding an organic solvent and a magnetic bead solution to the first mixed solution, oscillating and mixing, placing it on a magnetic separator until the solution is clarified, discarding the supernatant, and obtaining a first precipitate; S3, adding a first rinse solution to the first precipitate, oscillating and mixing, placing it on a magnetic separator until the solution is clarified, discarding the supernatant, and obtaining a second precipitate; adding a second rinse solution to the second precipitate, oscillating and mixing, placing it on a magnetic separator until the solution is clarified, discarding the supernatant, and obtaining a third precipitate; S4, drying the third precipitate, adding an eluent, oscillating and mixing, and then incubating, and then placing it on a magnetic separator until the solution is clarified, and the obtained supernatant is the blood genomic DNA.

[0016] In some embodiments, in step S1, the volume ratio of the blood sample to be extracted, the lysate and the proteinase K solution is (8-12): (0.5-1.5): (13-17), for example, it can be 8:0.5:13, 8:1:15, 8:1.5:17, 10:0.5:13, 10:1:15, 10:1.5:17, 12:0.5:13, 12:1:15, 12:1.5:17 or other ratios within this range; the incubation temperature is 60-70°C, for example, it can be 60°C, 62°C, 65°C, 68°C, 70°C or other values ​​within this range; the incubation time is 10-20 min, for example, it can be 10 min, 12 min, 15 min, 18 min, 20 min or other values ​​within this range.

[0017] In some embodiments, in step S2, the volume ratio of the organic solvent, the magnetic bead solution and the blood sample to be extracted is (1-2): (0.05-0.15): 1, for example, it can be 1:0.05:1, 1:0.1:1, 1:0.15:1, 1.5:0.05:1, 1.5:0.1:1, 1.5:0.15:1, 2:0.05:1, 2:0.1:1, 2:0.15:1 or other ratios within this range; wherein the organic solvent is selected from at least one of isopropanol and anhydrous ethanol, and the oscillation mixing time is 5-15 min, for example, it can be 5 min, 7 min, 10 min, 13 min, 15 min or other values ​​within this range.

[0018] In some embodiments, in step S3, the volume ratio of the first rinse liquid to the blood sample to be extracted is (2-3):1, for example, it can be 2:1, 2.2:1, 2.5:1, 2.8:1, 3:1 or other ratios within this range; the volume ratio of the second rinse liquid to the blood sample to be extracted is (2-4):1, for example, it can be 2:1, 2.5:1, 3:1, 3.5:1, 4:1 or other ratios within this range.

[0019] In some embodiments, in step S4, the drying temperature is 37-56°C, for example, 37°C, 40°C, 45°C, 50°C, 56°C or other values ​​within the range; the drying time is 3-5 min, for example, 3 min, 4 min, 5 min or other values ​​within the range; the volume ratio of the eluent to the blood sample to be extracted is (0.25-0.5):1, for example, 0.25:1, 0.3:1, 0.35:1, 0.4:1, 0.5:1 or other ratios within the range; the incubation temperature after oscillation mixing is 50-60°C, for example, 50°C, 52°C, 54°C, 56°C, 58°C, 60°C or other values ​​within the range; the time is 5-10 min, for example, 5 min, 6 min, 7 min, 8 min, 9 min, 10 min or other values ​​within the range.

[0020] The beneficial effect of the present invention is that, different from the prior art, the present invention optimizes the components of the lysate in the provided blood genomic DNA extraction kit, and by selecting specific contents of guanidine hydrochloride, NaCl, EDTA, dextran, Triton X-100, dithiothreitol, Tween-20, and trehalose as the components of the lysate, the above substances act synergistically, and at the same time, significantly improve the cell lysis efficiency, and can also obtain DNA molecules with higher purity, more complete structure and higher extraction yield. DETAILED DESCRIPTION

[0021] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0022] The experimental methods without specific conditions in the examples are usually carried out according to conventional conditions and conditions described in the manual, or according to conditions recommended by the manufacturer. The general equipment, materials, reagents, etc. used are all available from commercial channels unless otherwise specified.

[0023] Example 1 In this embodiment, the blood genomic DNA extraction kit includes a lysis solution, a first rinse solution, a second rinse solution, an elution solution, a proteinase K solution and a magnetic bead solution; The lysis solution includes the following components: 6M guanidine hydrochloride, 0.3M NaCl, 0.015M EDTA, 0.015M dextran, 2% (v / v) Triton X-100, 0.1M dithiothreitol, 10% (v / v) Tween-20, and 50mM trehalose; the first rinse solution includes the following components: 5M guanidine hydrochloride, 15mM Tris-HCl, 2mM EDTA, 0.3mM phenylmethylsulfonyl fluoride, and 25% (v / v) PEG 8000; the second rinse solution includes the following components: 0.25M Tris, 0.02M SDS, 0.5M NaCl, 0.06M EDTA, and 0.03% (v / v) sodium azide; the elution solution includes the following components: 10mM Tris-HCl, 2mM EDTA, pH 8.2; the concentration of proteinase K solution is 150 μg / mL, and the concentration of magnetic bead solution is 8 mg / mL.

[0024] The method for extracting blood genomic DNA using the extraction kit comprises the following steps: S1. Add 200 μL of the blood sample to be extracted into a centrifuge tube, then add 300 μL of lysis solution and 20 μL of proteinase K solution, shake and mix, and incubate at 65°C for 15 minutes. Shake and mix 2-3 times during the incubation period to obtain a first mixed solution; S2. Add 300 μL of isopropanol (or 400 μL of anhydrous ethanol) and 20 μL of magnetic bead solution to the first mixed solution. After oscillating and mixing for 10 minutes, place the centrifuge tube on a magnetic separator until the solution is clear, and discard the supernatant to obtain a first precipitate. S3, adding 500 μL of the first rinsing solution to the first precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a second precipitate; adding 600 μL of the second rinsing solution to the second precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a third precipitate; S4. Place the centrifuge tube containing the third precipitate in an oven at 40°C for 4 minutes, then add 80 μL of elution solution to the centrifuge tube. After oscillation and mixing, incubate in a 56°C water bath for 10 minutes. Then place the centrifuge tube on a magnetic separator until the solution is clarified. The resulting supernatant is the blood genomic DNA.

[0025] Example 2 In this embodiment, the blood genomic DNA extraction kit includes a lysis solution, a first rinse solution, a second rinse solution, an elution solution, a proteinase K solution and a magnetic bead solution; The lysate includes the following components: 5.8M guanidine hydrochloride, 0.25M NaCl, 0.013M EDTA, 0.013M dextran, 1.8% (v / v) Triton X-100, 0.08M dithiothreitol, 8% (v / v) Tween-20, and 13mM trehalose; the first rinse solution includes the following components: 4.8M guanidine hydrochloride, 13mM Tris-HCl, 1.5mM EDTA, 0.2mM phenylmethylsulfonyl fluoride, and 18% (v / v) PEG 8000; the second rinse solution includes the following components: 0.2M Tris, 0.015M SDS, 0.4M NaCl, 0.03M EDTA, and 0.02% (v / v) sodium azide; the elution solution includes the following components: 8mM Tris-HCl, 1.5mM EDTA, pH 7.8; the concentration of proteinase K solution is 120 μg / mL, and the concentration of magnetic bead solution is 5 mg / mL.

[0026] The method for extracting blood genomic DNA using the extraction kit comprises the following steps: S1. Add 200 μL of the blood sample to be extracted into a centrifuge tube, then add 300 μL of lysis solution and 20 μL of proteinase K solution, shake and mix, and incubate at 65°C for 15 minutes. Shake and mix 2-3 times during the incubation period to obtain a first mixed solution; S2. Add 300 μL of isopropanol (or 400 μL of anhydrous ethanol) and 20 μL of magnetic bead solution to the first mixed solution. After oscillating and mixing for 10 minutes, place the centrifuge tube on a magnetic separator until the solution is clear, and discard the supernatant to obtain a first precipitate. S3, adding 500 μL of the first rinsing solution to the first precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a second precipitate; adding 600 μL of the second rinsing solution to the second precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a third precipitate; S4. Place the centrifuge tube containing the third precipitate in an oven at 40°C for 4 minutes, then add 80 μL of elution solution to the centrifuge tube. After oscillation and mixing, incubate in a 56°C water bath for 10 minutes. Then place the centrifuge tube on a magnetic separator until the solution is clarified. The resulting supernatant is the blood genomic DNA.

[0027] Example 3 In this embodiment, the blood genomic DNA extraction kit includes a lysis solution, a first rinse solution, a second rinse solution, an elution solution, a proteinase K solution and a magnetic bead solution; The lysis solution includes the following components: 5.5M guanidine hydrochloride, 0.2M NaCl, 0.01M EDTA, 0.01M dextran, 1.5% (v / v) Triton X-100, 0.06M dithiothreitol, 5% (v / v) Tween-20, and 10mM trehalose; the first rinse solution includes the following components: 4.5M guanidine hydrochloride, 10mM Tris-HCl, 1mM EDTA, 0.1mM phenylmethylsulfonyl fluoride, and 10% (v / v) PEG 8000; the second rinse solution includes the following components: 0.15M Tris, 0.01M SDS, 0.3M NaCl, 0.005M EDTA, and 0.01% (v / v) sodium azide; the elution solution includes the following components: 3mM Tris-HCl, 1mM EDTA, pH 7.2; the concentration of proteinase K solution is 80 μg / mL, and the concentration of magnetic bead solution is 2 mg / mL.

[0028] The method for extracting blood genomic DNA using the extraction kit comprises the following steps: S1. Add 200 μL of the blood sample to be extracted into a centrifuge tube, then add 300 μL of lysis solution and 20 μL of proteinase K solution, shake and mix, and incubate at 65°C for 15 minutes. Shake and mix 2-3 times during the incubation period to obtain a first mixed solution; S2. Add 300 μL of isopropanol (or 400 μL of anhydrous ethanol) and 20 μL of magnetic bead solution to the first mixed solution. After oscillating and mixing for 10 minutes, place the centrifuge tube on a magnetic separator until the solution is clear, and discard the supernatant to obtain a first precipitate. S3, adding 500 μL of the first rinsing solution to the first precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a second precipitate; adding 600 μL of the second rinsing solution to the second precipitate, after oscillating and mixing, placing the centrifuge tube on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a third precipitate; S4. Place the centrifuge tube containing the third precipitate in an oven at 40°C for 4 minutes, then add 80 μL of elution solution to the centrifuge tube. After oscillation and mixing, incubate in a 56°C water bath for 10 minutes. Then place the centrifuge tube on a magnetic separator until the solution is clarified. The resulting supernatant is the blood genomic DNA.

[0029] Comparative Example 1 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that dithiothreitol is not added to the lysis solution.

[0030] Comparative Example 2 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that Tween-20 is not added to the lysate.

[0031] Comparative Example 3 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that no trehalose is added to the lysis solution.

[0032] Comparative Example 4 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that phenylmethylsulfonyl fluoride is not added to the first rinse solution.

[0033] Comparative Example 5 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that PEG 8000 is not added to the first rinse solution.

[0034] Comparative Example 6 In this comparative example, the blood genomic DNA extraction kit and the method for extracting blood genomic DNA using the extraction kit are basically the same as those in Example 1, except that sodium azide is not added to the second rinse solution.

[0035] Performance Testing The concentration and purity of the blood genomic DNA extracted in Examples 1-3 and Comparative Examples 1-6 were tested, and the results are shown in Table 1 below.

[0036] Table 1 Concentration and purity of blood genomic DNA

[0037] It can be seen from the data in Table 1 that the blood genomic DNA extracted by using the kit in Examples 1-3 has good concentration and purity; the lysate in the kit in Comparative Examples 1-3 does not add dithiothreitol, Tween-20 and trehalose, and it is found that the concentration and purity of the extracted blood genomic DNA are both reduced to a certain extent; the first rinse solution in the kit in Comparative Examples 4-5 does not add phenylmethylsulfonyl fluoride and PEG 8000, and the second rinse solution in the kit in Comparative Example 6 does not add sodium azide. It is found that the concentration and purity of the extracted blood genomic DNA are both reduced to a certain extent; the above results show that the components in the blood genomic DNA extraction kit provided by the present invention have a synergistic effect, which can improve the concentration and purity of the extracted blood genomic DNA.

[0038] In summary, the present invention optimizes the components of the lysate in the provided blood genomic DNA extraction kit. Through the synergistic effect of the above components, while significantly improving the cell lysis efficiency, it can also obtain DNA molecules with higher purity, more complete structure and higher extraction yield.

[0039] It should be noted that the above embodiments all belong to the same inventive concept, and the description of each embodiment has its own focus. For matters that are not described in detail in some embodiments, reference may be made to the description in other embodiments.

[0040] The above-mentioned embodiments only express the implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A blood genomic DNA extraction kit, characterized in that: comprising a lysing solution, a first rinsing solution, a second rinsing solution and an eluting solution; The lysate comprises the following components: 5.5-6M guanidine hydrochloride, 0.2-0.3M NaCl, 0.01-0.015M EDTA, 0.01-0.015M dextran, 1.5-2% Triton X-100, 0.06-0.1M dithiothreitol, 5-10% Tween-20, and 10-50mM trehalose.

2. The blood genomic DNA extraction kit according to claim 1, characterized in that: The first washing solution comprises the following components: 4.5-5M guanidine hydrochloride, 10-15mM Tris-HCl, 1-2mM EDTA, 0.1-0.3mM phenylmethylsulfonyl fluoride, and PEG 8000 with a volume concentration of 10-25%.

3. The blood genomic DNA extraction kit according to claim 1, characterized in that: The second rinsing solution includes the following components: 0.15-0.25M Tris, 0.01-0.02M SDS, 0.3-0.5M NaCl, 0.005-0.06M EDTA, and sodium azide with a volume concentration of 0.01-0.03%.

4. The blood genomic DNA extraction kit according to claim 1, characterized in that: The eluent comprises the following components: 3-10 mM Tris-HCl, 1-2 mM EDTA, pH 7.2-8.

2.

5. The blood genomic DNA extraction kit according to claim 1, characterized in that: The blood genomic DNA extraction kit also includes a proteinase K solution and a magnetic bead solution; Wherein, the concentration of the proteinase K solution is 80-150 μg / mL, and the concentration of the magnetic bead solution is 2-8 mg / mL.

6. A method for extracting blood genomic DNA using the extraction kit according to any one of claims 1 to 5, characterized in that: The steps include: S1, adding a lysate and a proteinase K solution to the blood sample to be extracted, shaking and mixing, and then incubating to obtain a first mixed solution; S2, adding an organic solvent and a magnetic bead solution to the first mixed solution, shaking and mixing, placing the mixture on a magnetic separator until the solution is clear, and discarding the supernatant to obtain a first precipitate; S3, adding the first rinsing liquid to the first precipitate, after oscillating and mixing, placing it on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a second precipitate; adding the second rinsing liquid to the second precipitate, after oscillating and mixing, placing it on a magnetic separator until the solution is clear, discarding the supernatant, and obtaining a third precipitate; S4. Dry the third precipitate, add the eluent, incubate after oscillation mixing, and then place it on a magnetic separator until the solution is clarified. The obtained supernatant is the blood genomic DNA.

7. The method according to claim 6, characterized in that In step S1, the volume ratio of the blood sample to be extracted, the lysate and the proteinase K solution is (8-12): (0.5-1.5): (13-17), the incubation temperature is 60-70°C, and the incubation time is 10-20 minutes.

8. The method according to claim 6, characterized in that In step S2, the volume ratio of the organic solvent, the magnetic bead solution and the blood sample to be extracted is (1-2): (0.05-0.15): 1; Wherein, the organic solvent is selected from at least one of isopropanol and anhydrous ethanol, and the oscillation mixing time is 5-15 minutes.

9. The method according to claim 6, characterized in that In step S3, the volume ratio of the first rinsing liquid to the blood sample to be extracted is (2-3):1, and the volume ratio of the second rinsing liquid to the blood sample to be extracted is (2-4):

1.

10. The method according to claim 6, characterized in that In step S4, the drying temperature is 37-56°C and the time is 3-5 minutes; the volume ratio of the eluent to the blood sample to be extracted is (0.25-0.5):1, and the incubation temperature after oscillation mixing is 50-60°C and the time is 5-10 minutes.

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