Lysate for nucleic acid extraction and application thereof

Abstract

The invention discloses a lysate for nucleic acid extraction and application thereof, and relates to the technical field of molecular biology. The lysate is prepared from the following components with final concentration: 20 to 150mM of alkaline substance, 0.5 to 5M of penetrant, 0.05 to 5.5 w / v percent of compound surfactant and 0.01 to 1mg / mL of protease K; the compound surfactant comprises nonylphenol polyoxyethylene ether and lauryl sodium sulfate. The lysate can efficiently lyse animal tissue cells, release high-quality genome DNA (deoxyribonucleic acid), provide sufficient templates for PCR (polymerase chain reaction) amplification and meet the requirement of direct PCR amplification; components in the lysate are stable, and the lysate is easy to store and transport and suitable for large-scale sample detection. In addition, the lysis buffer is suitable for lysis and PCR amplification of various animal tissue samples, and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of molecular biology, and in particular to a lysate for nucleic acid extraction and an application thereof. Background Art

[0002] In the fields of biology, medicine, and agricultural science, PCR amplification technology is an important molecular zoology tool used to detect and analyze DNA sequences in animal samples. However, the traditional PCR amplification method requires tissue lysis and DNA extraction of the sample first. These steps are not only time-consuming and labor-intensive, but may also cause sample loss, contamination, or degradation, thereby affecting the effect and accuracy of PCR amplification. Therefore, the development of a lysis solution that can efficiently lyse animal tissue cells and directly use it for PCR amplification is of great significance for simplifying experimental steps and improving detection efficiency and accuracy.

[0003] In view of this, the present invention is proposed. Summary of the invention

[0004] The object of the present invention is to provide a lysis solution for nucleic acid extraction and application thereof, so as to solve the above technical problems.

[0005] The present invention is achieved in that:

[0006] In a first aspect, an embodiment of the present invention provides a lysate for nucleic acid extraction, comprising the following components at final concentrations:

[0007] 20mM-150mM alkaline substance, 0.5M-5M osmotic agent, 0.05w / v%-5.5w / v% complex surfactant and 0.01mg / mL-1mg / mL proteinase K;

[0008] The compound surfactant includes nonylphenol polyoxyethylene ether and sodium lauryl sulfate.

[0009] In a second aspect, an embodiment of the present invention provides a use of the aforementioned lysis solution in nucleic acid extraction or PCR amplification.

[0010] In a third aspect, an embodiment of the present invention provides a reagent or a kit, which includes the aforementioned lysis solution.

[0011] In a fourth aspect, an embodiment of the present invention provides a method for extracting nucleic acid, comprising contacting a sample with the aforementioned lysis solution; and incubating the sample at a temperature of 50° C.-55° C. for 10 min-30 min.

[0012] In a fifth aspect, an embodiment of the present invention provides a method for nucleic acid detection, wherein a sample is contacted with the aforementioned lysis solution; and a nucleic acid extract is taken for nucleic acid amplification.

[0013] The present invention has the following beneficial effects:

[0014] The lysing solution for nucleic acid extraction provided by the present invention can efficiently lyse animal tissue cells, release high-quality genomic DNA, provide sufficient templates for PCR amplification, and meet the needs of direct PCR amplification; the components in the lysing solution are stable, easy to store and transport, and suitable for large-scale sample detection. In addition, the lysing solution for nucleic acid extraction provided by the present invention is suitable for lysis and PCR amplification of various animal tissue samples, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0016] Figure 1 This is the result of mouse ear lysis amplification;

[0017] Figure 2 This is the result of mouse toe cleavage amplification;

[0018] Figure 3 The results of lysis and amplification of different mouse tissues;

[0019] Figure 4 This is the result of suspension cell lysis and amplification;

[0020] Figure 5 The results of screening for sodium hydroxide in the lysate;

[0021] Figure 6 The results of SDS screening in lysate;

[0022] Figure 7 The results are for the screening of proteinase K in the lysate;

[0023] Figure 8 The results of screening for betaine in the lysate are shown in Figure 2.

[0024] Fig. 9 These are the screening results of nonylphenol polyoxyethylene ether in the lysate. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0026] In a first aspect, an embodiment of the present invention provides a lysate for nucleic acid extraction, comprising the following components at final concentrations:

[0027] 20mM-150mM alkaline substance, 0.5M-5M osmotic agent, 0.05w / v%-5.5w / v% complex surfactant and 0.01mg / mL-1mg / mL proteinase K;

[0028] The compound surfactant includes nonylphenol polyoxyethylene ether and sodium lauryl sulfate.

[0029] It should be noted that the lysis solution provided in the embodiment of the present invention has a highly efficient cell destruction ability and can achieve precise DNA release; the components can quickly penetrate into the interior of the cell, destroy the cell membrane structure through a specific chemical mechanism, and thereby promote the effective release of DNA molecules into the lysis solution.

[0030] Surfactants have excellent wettability and dispersibility. The present invention uses a compound surfactant to have a synergistic effect, which effectively promotes cell lysis and the release of DNA, while reducing the interference of cell debris and impurities, thereby ensuring that the purity of the extracted DNA reaches a high level. In addition, the compound surfactant can reduce the usage of a single surfactant, thereby reducing overall costs. By selecting a suitable compatibility, the use effect of the compound surfactant is usually better than that of a single surfactant, thereby reducing the usage of the surfactant, reducing costs, and improving efficiency of use.

[0031] In an optional embodiment of the present invention, the final concentration of the alkaline substance is 50 mM-80 mM, which can be selected from any one of 50 mM, 55 mM, 64 mM, 70 mM and 80 mM, or other values ​​within the range of 50 mM-80 mM.

[0032] It should be noted that alkaline substances can effectively destroy cell structures by adjusting the pH value of the solution. Alkaline components within the set range can effectively destroy cell structures while avoiding unnecessary damage to DNA, finding the best balance between cell lysis effect and DNA stability.

[0033] Furthermore, the alkaline substance is selected from at least one of sodium hydroxide, potassium hydroxide and lithium hydroxide, preferably sodium hydroxide. In other embodiments of the present invention, the alkaline substance can be selected from potassium hydroxide or lithium hydroxide according to actual needs.

[0034] In an optional embodiment of the present invention, the final concentration of the osmotic agent is 1 M-3 M. It can be selected from any one of 1 M, 1.8 M, 2 M and 3 M, or other values ​​within the range of 1 M-3 M.

[0035] It should be noted that osmotic agents are key components for regulating osmotic pressure, maintaining DNA stability, and improving DNA solubility. By regulating the osmotic pressure of the solution, it can effectively promote cell lysis and DNA release, while protecting DNA from physical and chemical damage.

[0036] Furthermore, the osmotic agent is selected from at least one of betaine, potassium chloride, sodium chloride, and sucrose, preferably betaine. In other embodiments of the present invention, the osmotic agent can be selected from at least one of potassium chloride, sodium chloride, or sucrose according to actual needs.

[0037] In an optional embodiment of the present invention, the final concentration of the compound surfactant is 1 w / v%-5.5 w / v%.

[0038] In an optional embodiment of the present invention, in the compound surfactant, the final concentration of nonylphenol polyoxyethylene ether is 0.1w / v%-1w / v%, and the final concentration of sodium dodecyl sulfate (SDS) is 1w / v%-5w / v%. Wherein, the final concentration of nonylphenol polyoxyethylene ether can be selected from any one of 0.1w / v%, 0.2w / v%, 0.5w / v%, 0.8w / v% and 1w / v%, or other values ​​within the range of 0.1w / v%-1w / v%; the final concentration of sodium dodecyl sulfate can be selected from any one of 1w / v%, 2w / v%, 3w / v%, 4.0w / v% and 5w / v%, or other values ​​within the range of 1w / v%-5w / v%.

[0039] Furthermore, the compound surfactant is 0.5% nonylphenol polyoxyethylene ether and 3% sodium dodecyl sulfate. At this time, the cell lysis effect and DNA purity of the lysate have reached the best state.

[0040] In an optional embodiment of the present invention, the final concentration of proteinase K is 0.1 mg / mL-1 mg / mL.

[0041] It should be noted that proteinase K can specifically and efficiently degrade proteins in cells, allowing DNA to be fully released without causing any damage to the DNA.

[0042] Further, the final concentration of proteinase K is 0.4 mg / mL-0.6 mg / mL, which can be selected from any one of 0.4 mg / mL, 0.5 mg / mL, 0.55 mg / mL and 0.6 mg / mL, or other values ​​within the range of 0.4 mg / mL-0.6 mg / mL.

[0043] In a second aspect, an embodiment of the present invention provides a use of the aforementioned lysis solution in nucleic acid extraction or PCR amplification.

[0044] In a third aspect, an embodiment of the present invention provides a reagent or a kit, which includes the lysis solution as described above.

[0045] In a fourth aspect, an embodiment of the present invention provides a method for extracting nucleic acid, comprising contacting a sample with the aforementioned lysis solution; and incubating the sample at a temperature of 50° C.-55° C. for 10 min-30 min.

[0046] In a fifth aspect, an embodiment of the present invention provides a method for nucleic acid detection, wherein a sample is contacted with the aforementioned lysis solution; and a nucleic acid extract is taken for nucleic acid amplification.

[0047] In an optional embodiment of the present invention, when the lysate is used in PCR amplification, the steps include:

[0048] First, a lysis solution is prepared, which includes the following steps: alkaline substances, osmotic agents, compound surfactants and water are mixed in proportion, and stirred evenly until completely dissolved; then proteinase K is added until completely dissolved, and the pH value of the system is adjusted to 7.5-10 to obtain a lysis solution for nucleic acid extraction.

[0049] It should be noted that the raw materials used in the preparation of the lysate are selected to meet the biological experiment grade or higher purity requirements; a high-precision electronic balance is used to accurately weigh each raw material. The water used in the preparation of the lysate can be selected from deionized water, ultrapure water, etc. according to actual needs.

[0050] The addition of proteinase K requires controlling the temperature of the system to avoid enzyme inactivation due to excessive temperature. The specific temperature is set according to the requirements of proteinase K. The proteinase K used in the embodiments of the present invention is derived from Sangon Biotech (Shanghai) Co., Ltd. (Article No. B600169).

[0051] In an optional embodiment of the present invention, during the preparation of the lysate, after the pH is adjusted, the lysate is filtered using a sterile filter; it is dispensed into sterile containers, sealed and labeled with relevant information, such as preparation time, preparation personnel, storage temperature and expiration date, etc. It is then stored at a specific storage temperature, avoiding direct sunlight and high temperature environments.

[0052] It should be noted that after the lysate is prepared, an appearance inspection is required to ensure that the lysate is clear and transparent; the pH value is rechecked to ensure that it is within the target range of 7.5-10.

[0053] The lysis solution is used in nucleic acid extraction or PCR amplification, specifically, it also includes sample preparation, lysis treatment, incubation and inactivation, and PCR amplification treatment.

[0054] Sample preparation: Take fresh or frozen animal tissue or cell samples and cut them precisely using sterile surgical scissors or blades to ensure consistent sample size.

[0055] It should be noted that the research of the present invention is not for the purpose of disease diagnosis, and the animal tissues used in the experiment can be selected from the ears, liver, tail and toes of animals according to actual needs. In the embodiments of the present invention, mouse-related tissues are used.

[0056] Lysis treatment: Transfer the cut animal tissue samples to a PCR tube or a 1.5mL centrifuge tube, then add 10μL-200μL of lysis solution to the tube, so that the animal tissue samples are in contact with the lysis solution or completely immersed in the lysis solution.

[0057] Incubation and inactivation: Place the PCR tube or centrifuge tube containing the animal tissue sample and lysate in a PCR instrument or metal bath, set the temperature to 50℃-55℃, and incubate for 10min-30min to allow the lysate to fully act, destroy the tissue cell structure, and release DNA. After that, raise the temperature to 90℃-98℃ and maintain it for 5min-10min to inactivate the enzyme in the lysate to prevent interference with subsequent PCR amplification.

[0058] It should be noted that the specific incubation instrument used can be a PCR tube or a centrifuge tube according to actual needs.

[0059] PCR amplification: Take the lysed product as a template and add it directly to the PCR system to amplify according to the preset PCR amplification program. After amplification, detect the PCR product to evaluate the effect of the lysate.

[0060] It should be noted that the PCR reagent used in the embodiment of the present invention is 2×PCR mix, which is derived from Sangon Biotech (Shanghai) Co., Ltd. (specifically named 2X HyperMB Taq PCR Master Mix Pro).

[0061] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0062] Example 1

[0063] This embodiment provides a lysis solution for nucleic acid extraction, and the preparation method thereof includes the following steps:

[0064] (1) Weigh 0.28 g NaOH, 23.43 g betaine, 0.5 g nonylphenol polyoxyethylene ether and 3 g sodium dodecyl sulfate, add deionized water, stir evenly until completely dissolved, and dilute to 100 mL with deionized water.

[0065] (2) Add 50 mg of proteinase K to the solution obtained in (1) at 25°C and continue stirring until completely mixed.

[0066] (3) Using a pH meter, adjust the pH of the solution obtained in (2) to 8.0 to obtain a lysate for nucleic acid extraction.

[0067] Experimental Example 1

[0068] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, as follows:

[0069] (1) Sample preparation: Use sterile surgical scissors or a blade to precisely cut fresh or frozen 2 mm 2 Mouse ear tissue, set aside.

[0070] (2) Lysis treatment: The cut mouse ear tissue sample was transferred to a 1.5 mL centrifuge tube, and then 100 μL of the lysis solution prepared in Example 1 was added to the centrifuge tube to ensure that the mouse ear tissue sample was completely immersed therein.

[0071] (3) Incubation and inactivation: Place the centrifuge tube containing the mouse ear tissue sample and lysate in a metal bath, set the temperature to 50°C, and incubate for 30 minutes to allow the lysate to fully act, destroy the tissue cell structure, and release DNA. Then, raise the temperature to 95°C and maintain it for 5 minutes to inactivate the proteinase K in the lysate to prevent it from interfering with subsequent PCR amplification.

[0072] (4) PCR amplification: Take 1 μL of the lysate as a template and directly add it into the PCR system for amplification. The PCR system is shown in Table 1. After mixing, run the amplification program shown in Table 2 on the PCR instrument.

[0073] Table 1 PCR system

[0074] Components Addition amount / μl Cleavage products 1 Upstream primer (10 μM) 2 Downstream primer (10 μM) 2 Double distilled water 20 2×PCR Mix 25 Total volume 50

[0075] Table 2 Amplification procedures

[0076]

[0077] Experimental Example 2

[0078] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 only in that:

[0079] (1) Sample preparation: Use sterile surgical scissors or a blade to precisely cut a fresh or frozen mouse toe (2 mm in length) and set aside.

[0080] Experimental Example 3

[0081] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 only in that:

[0082] (1) Sample preparation: Use sterile surgical scissors or a blade to precisely cut fresh or frozen 2 mm mouse tail tissue and set aside.

[0083] Experimental Example 4

[0084] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 only in that:

[0085] (1) Sample preparation: Use sterile surgical scissors or a blade to precisely cut fresh or frozen 1 mm 3 Mouse muscle, reserved.

[0086] Experimental Example 5

[0087] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 only in that:

[0088] (1) Sample preparation: Take 2 μL of cell suspension (9000 suspended cells), collect and count using sterile methods.

[0089] (2) Lysis treatment: According to the number of cells, add 50 μL of the lysis solution prepared in Example 1 to the cell sample to ensure that the cells are completely immersed therein. Gently blow or vortex to mix to promote cell lysis.

[0090] (3) Incubation and inactivation: Place the mixture containing the cell sample and lysis solution in a metal bath, set the temperature to 50°C, and incubate for 30 min to allow the lysis solution to fully act and destroy the suspended cell structure; then, raise the temperature to 95°C and maintain it for 5 min to inactivate the proteinase K in the lysis solution to prevent it from interfering with subsequent PCR amplification.

[0091] (4) PCR amplification: Take 1 μL of the obtained lysis product as a template and directly add it into the PCR system for PCR amplification. The PCR system is shown in Table 1, and the amplification program is shown in Table 2.

[0092] Test Example 1

[0093] This test example studies the final concentration of sodium hydroxide (NaOH) in the lysis solution used for nucleic acid extraction, and the specific implementation process is as follows:

[0094] (1) Weigh a gradient mass of NaOH, 23.43 g of betaine, 0.5 g of nonylphenol polyoxyethylene ether, and 3 g of sodium dodecyl sulfate, add deionized water, stir evenly until completely dissolved, and adjust the volume to 100 mL.

[0095] The gradient masses of sodium hydroxide are 0.12 g, 0.2 g, 0.28 g, 0.4 g, and 0.6 g, and the corresponding molar concentrations are 30 mM, 50 mM, 70 mM, 100 mM, and 150 mM, respectively.

[0096] (2) Add 50 mg of proteinase K to the solution obtained in (1) at 25°C and continue stirring until it is completely dissolved.

[0097] (3) Using a pH meter, adjust the pH of the solution obtained in (2) to 8.0 to obtain a lysate for nucleic acid extraction.

[0098] (4) Use the lysis solution obtained in (3) to lyse a mouse toe (length 2 mm). The lysis process is as follows: add 50 μL of lysis solution to a 200 μL PCR tube, then add the obtained mouse toe thereto, set the temperature at 50°C in a metal bath and incubate for 30 min. Take 1 μL of the obtained lysis product as a template and add it to the PCR amplification system. Run the amplification program for amplification. The PCR system is shown in Table 1, and the amplification program is shown in Table 2.

[0099] Test Example 2

[0100] This test example studies the final concentration of sodium dodecyl sulfate (SDS) in the lysis solution used for nucleic acid extraction. The specific implementation process is different from that of Test Example 1 only in that:

[0101] (1) Weigh 0.28 g NaOH, 23.43 g betaine, 0.5 g nonylphenol polyoxyethylene ether and gradient mass sodium dodecyl sulfate, add deionized water, stir evenly until completely dissolved, and dilute to 100 mL with deionized water.

[0102] The gradient masses of SDS are 1 g, 2 g, 3 g, 4 g, and 5 g, and the corresponding mass concentrations are 1%, 2%, 3%, 4%, and 5%, respectively.

[0103] Test Example 3

[0104] This test example studies the final concentration of proteinase K in the lysis solution used for nucleic acid extraction. The specific implementation process is different from that of Test Example 1 only in that:

[0105] (2) Add gradient amounts of proteinase K to the solution obtained in (1) at 25°C and continue stirring until it is completely dissolved.

[0106] Among them, the gradient mass of proteinase K is 10 mg, 30 mg, 50 mg, 70 mg, and 100 mg, and the corresponding mass concentrations are 0.1 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.7 mg / mL, and 1 mg / mL, respectively.

[0107] Test Example 4

[0108] This test example studies the final concentration of betaine in the lysate for nucleic acid extraction. The specific implementation process is different from that of Test Example 1 only in that:

[0109] (1) Weigh 0.28 g NaOH, gradient betaine, 0.5 g nonylphenol polyoxyethylene ether and 3 g sodium dodecyl sulfate, add deionized water, stir evenly until completely dissolved, and dilute to 100 mL with deionized water.

[0110] The gradient masses of betaine are 11.175 g, 23.43 g, 34.145 g, 46.86 g, and 58.575 g, and the corresponding molar concentrations are 1 M, 2 M, 3 M, 4 M, and 5 M, respectively.

[0111] Test Example 5

[0112] This test example studies the final concentration of nonylphenol polyoxyethylene ether in the lysate for nucleic acid extraction. The specific implementation process is different from that of Test Example 1 only in that:

[0113] (1) Weigh 0.28 g NaOH, 23.43 g betaine, gradient mass nonylphenol polyoxyethylene ether and 3 g sodium dodecyl sulfate, add deionized water, stir evenly until completely dissolved, and dilute to 100 mL with deionized water.

[0114] The gradient masses of nonylphenol polyoxyethylene ether are 0.2g, 0.5g, 0.8g, and 1g, and the corresponding mass concentrations are 0.2%, 0.5%, 0.8%, and 1%, respectively.

[0115] Test Example 6

[0116] This test example detects the PCR amplification products in Experimental Examples 1-5 and Experimental Examples 1-5, and specifically evaluates the performance of the lysate by electrophoresis. The detection method is as follows: 0.8% agarose gel is prepared using 1×TAE buffer, the sample volume is 5μL, and electrophoresis is performed under the conditions of 180V 15min (Junyi JY600C electrophoresis instrument power supply and matching horizontal electrophoresis tank), and then ultraviolet transmission light exposure is performed for 200ms (Bolu Teng GelView 5000ProⅡ) to obtain gel imaging results.

[0117] The test results are as follows:

[0118] Figure 1 This is the result of mouse ear lysis amplification; Figure 2 This is the result of mouse toe cleavage amplification; Figure 3 The results of lysis and amplification of different mouse tissues; Figure 4 The results are for suspension cell lysis and amplification.

[0119] from Figure 1-Figure 4 It can be seen from the results that the lysis solution for nucleic acid extraction provided by the present invention can quickly and effectively lyse different types of animal tissue samples (ear tissue, muscle tissue, tail tissue and toes of mice) and cell samples, while maintaining the integrity of DNA, which is convenient for subsequent PCR amplification.

[0120] Figure 5 The results of screening for sodium hydroxide in the lysate; Figure 6 The results of SDS screening in lysate; Figure 7 The results are for the screening of proteinase K in the lysate; Figure 8 The results of screening for betaine in the lysate are shown in Figure 2. Fig. 9 These are the screening results of nonylphenol polyoxyethylene ether in the lysate.

[0121] from Figure 5-Figure 9 It can be clearly seen from the results that the core components of the lysate for nucleic acid extraction involved in the present invention are sodium hydroxide (or alkaline substances such as potassium hydroxide), SDS, proteinase K, and betaine, and nonylphenol polyoxyethylene ether mainly plays a promoting role in enhancing the effect.

[0122] In summary, the lysate for nucleic acid extraction provided by the present invention can release high-quality genomic DNA, provide sufficient templates for PCR amplification, and can quickly and effectively lyse different types of animal tissue samples and cell samples while maintaining the integrity of DNA; during PCR amplification, there is no need to perform cumbersome DNA extraction and purification steps, and the supernatant is directly taken for PCR amplification, which greatly improves the detection efficiency and accuracy; the components in the lysate are stable, easy to store and transport, and suitable for large-scale sample detection. The lysate of the present invention shows universality and stability in different sample types, which provides strong support for its wide application in the fields of molecular biology research, genetic diagnosis, genetic breeding, etc.

[0123] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A lysis solution for nucleic acid extraction, characterized in that: The lysate includes the following components at final concentrations: 20mM-150mM alkaline substance, 0.5M-5M osmotic agent, 0.05w / v%-5.5w / v% complex surfactant and 0.01mg / mL-1mg / mL proteinase K; The compound surfactant comprises nonylphenol polyoxyethylene ether and sodium lauryl sulfate.

2. The lysate according to claim 1, characterized in that The final concentration of the alkaline substance is 50mM-80mM; Preferably, the alkaline substance is selected from at least one of sodium hydroxide, potassium hydroxide and lithium hydroxide; preferably sodium hydroxide.

3. The lysate according to claim 1, characterized in that The final concentration of the osmotic agent is 1M-3M; Preferably, the osmotic agent is selected from at least one of betaine, potassium chloride, sodium chloride and sucrose; preferably betaine.

4. The lysate according to claim 1, characterized in that The final concentration of the compound surfactant is 1 w / v%-5.5 w / v%.

5. The lysate according to claim 4, characterized in that The final concentration of nonylphenol polyoxyethylene ether in the composite surfactant is 0.1 w / v%-1 w / v%, and the final concentration of sodium dodecyl sulfate is 1 w / v%-5 w / v%.

6. The lysate according to claim 1, characterized in that The final concentration of the proteinase K is 0.1 mg / mL-1 mg / mL.

7. Use of the lysate as claimed in claim 1 in nucleic acid extraction or PCR amplification.

8. A reagent or a kit, characterized in that: The reagent or kit comprises the lysate as claimed in claim 1.

9. A method for extracting nucleic acid, characterized in that: contacting the sample with the lysis solution as claimed in claim 1; Incubate at 50°C-55°C for 10 min-30 min.

10. A method for nucleic acid detection, characterized in that: The sample is contacted with the lysis solution as claimed in claim 1; and the nucleic acid extract is taken for nucleic acid amplification.

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