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 in final concentration: 10 to 100 mM of alkaline substances, 0.05 to 6 w / v percent of a compound surfactant, 0.1 to 0.5 mg / mL of protease K and a compound penetrant, wherein the concentration of the alkaline substances is 10 to 100 mM; the concentration of the compound surfactant is 0.05 to 6 w / v percent; the concentration of the protease K is 0.1 to 0.5 mg / mL. The preparation is simple, and during use, only a small amount of sample needs to be added into the lysate, so that the PCR amplification requirement can be met; the lysate can rapidly and efficiently lyse a sample and release a large amount of high-quality DNA, has good compatibility with a PCR amplification system, and can be directly used for PCR amplification to obtain a clear and specific amplification product.

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 application thereof. Background Art

[0002] In plant molecular biology research, polymerase chain reaction (PCR) is a commonly used technical means to amplify specific DNA fragments for gene analysis, genetic breeding, etc. However, traditional PCR amplification methods usually require complex DNA extraction and purification processes from plant tissues, which is not only time-consuming and labor-intensive, but also easily introduces contamination, affecting the efficiency and accuracy of PCR amplification.

[0003] In recent years, with the continuous development of molecular biology technology, some researchers have begun to try to directly lyse plant tissues for PCR amplification, the so-called "direct amplification" technology. This technology can greatly simplify the experimental steps and improve experimental efficiency. However, the existing lysis buffer still has shortcomings in lysis efficiency, DNA release amount and PCR amplification compatibility, which limits its wide application in plant tissue PCR direct amplification.

[0004] Therefore, developing an efficient, rapid and compatible plant tissue PCR direct amplification lysis solution is of great significance for promoting the development of plant molecular biology research.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] 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.

[0007] The present invention is achieved in that:

[0008] In a first aspect, an embodiment of the present invention provides a lysate for nucleic acid extraction, comprising the following final concentration components:

[0009] 10mM-100mM alkaline substance, 0.05w / v%-6w / v% compound surfactant, 0.1mg / mL-0.5mg / mL proteinase K and compound osmotic agent;

[0010] Wherein, the compound surfactant includes nonylphenol polyoxyethylene ether and sodium dodecyl sulfate;

[0011] The compound penetrant includes 1M-4M of penetrant A and 5w / v%-15w / v% of penetrant B.

[0012] 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.

[0013] In a third aspect, an embodiment of the present invention provides a reagent or a kit, which includes the aforementioned lysis solution.

[0014] 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.

[0015] 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.

[0016] The present invention has the following beneficial effects:

[0017] The lysate for nucleic acid extraction provided by the embodiment of the present invention is simple to prepare. When used, only a small amount of sample needs to be added to the lysate to meet the PCR amplification requirements. The sample can be lysed quickly and efficiently to release a large amount of high-quality DNA. The lysate has good compatibility with the PCR amplification system and can be directly used for PCR amplification to obtain clear and specific amplification products. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] 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.

[0019] Figure 1 This is the result of Arabidopsis leaf lysis amplification;

[0020] Figure 2 This is the result of rice leaf lysis amplification;

[0021] Figure 3 This is the result of maize leaf lysis amplification;

[0022] Figure 4 The results of screening for LiOH in the lysate;

[0023] Figure 5 The results of SDS screening in lysate;

[0024] Figure 6 The results are for the screening of proteinase K in the lysate;

[0025] Figure 7 The results of screening for betaine in the lysate are shown in Figure 2.

[0026] Figure 8The results are the screening results of nonylphenol polyoxyethylene ether in the pyrolysis solution;

[0027] Fig. 9 The results are shown for the screening of polysucrose-70 in the lysate. DETAILED DESCRIPTION

[0028] 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.

[0029] In a first aspect, an embodiment of the present invention provides a lysate for nucleic acid extraction, comprising the following components at final concentrations:

[0030] 10mM-100mM alkaline substance, 0.05w / v%-6w / v% compound surfactant, 0.1mg / mL-0.5mg / mL proteinase K and compound osmotic agent;

[0031] Wherein, the compound surfactant includes nonylphenol polyoxyethylene ether and sodium dodecyl sulfate;

[0032] The compound penetrant includes 1M-4M of penetrant A and 5w / v%-15w / v% of penetrant B.

[0033] It should be noted that the lysis solution for nucleic acid extraction provided in the embodiment of the present invention has a high efficiency in cell destruction and can achieve precise release of DNA; the components can quickly penetrate into the cell, destroy the cell wall and cell membrane structure through a specific chemical mechanism, and thus promote the effective release of DNA molecules into the lysis solution.

[0034] 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.

[0035] Osmotic agent is a key component for regulating osmotic pressure, maintaining DNA stability and improving DNA solubility. By adjusting the osmotic pressure of the solution, cell lysis and DNA release can be effectively promoted, while protecting DNA from physical and chemical damage. The present invention uses a compound of osmotic agent A and osmotic agent B, which has good compatibility, and synergistically promotes cell lysis without affecting the lysis effect, making cell lysis more thorough.

[0036] 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.

[0037] 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.

[0038] Furthermore, the alkaline substance is selected from at least one of sodium hydroxide, potassium hydroxide and lithium hydroxide; preferably lithium hydroxide.

[0039] In an optional embodiment of the present invention, the final concentration of the osmotic agent A is 1M-3M; it can be selected from any one of 1M, 1.5M, 2M and 3M, or other values ​​within the range of 1M-3M.

[0040] Furthermore, the osmotic agent A is selected from at least one of betaine, potassium chloride and sodium chloride; preferably betaine.

[0041] Further, the final concentration of osmotic agent B is 7w / v%-13w / v%, and osmotic agent B is polysucrose 70. It can be selected from any one of 7w / v%, 8w / v%, 10w / v% and 13w / v%, or other values ​​within the range of 7w / v%-13w / v%.

[0042] In an optional embodiment of the present invention, the final concentration of nonylphenol polyoxyethylene ether in the compound surfactant is 0.05w / v%-0.2w / v%, which can be selected from any one of 0.05w / v%, 0.06w / v%, 0.1w / v%, 0.15w / v% and 0.2w / v%, or other values ​​within the range of 0.05w / v%-0.2w / v%.

[0043] The final concentration of sodium dodecyl sulfate (SDS) is 1.5 w / v%-2.5 w / v%, which can be selected from any one of 1.5 w / v%, 1.6 w / v%, 1.8 w / v%, 2 w / v% and 2.5 w / v%, or other values ​​within the range of 1.5 w / v%-2.5 w / v%.

[0044] In an optional embodiment of the present invention, the final concentration of proteinase K is 0.2 mg / mL-0.4 mg / mL, which can be selected from any one of 0.2 mg / mL, 0.25 mg / mL, 0.3 mg / mL and 0.4 mg / mL, or other values ​​within the range of 0.2 mg / mL-0.4 mg / mL.

[0045] 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.

[0046] 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.

[0047] In a third aspect, an embodiment of the present invention provides a reagent or a kit, which includes the aforementioned lysis solution.

[0048] 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.

[0049] 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.

[0050] In an optional embodiment of the present invention, when the lysate for nucleic acid extraction is used in PCR amplification, the steps include:

[0051] First, the preparation of a lysate for nucleic acid extraction comprises the following steps:

[0052] (1) Prepare an appropriate amount of deionized water in a container and set aside.

[0053] It should be noted that the container is mainly used for the configuration of the lysis solution, and it can be selected according to actual needs, such as a beaker.

[0054] (2) Add alkaline substances, penetrant A, compound surfactants and penetrant B. Add each substance until it is completely dissolved before adding another substance to ensure that the substances are completely dissolved and the system is uniform.

[0055] (3) Finally, add proteinase K and stir thoroughly again.

[0056] (4) Add deionized water to the final volume and check the pH value (8-10) and other physical and chemical properties to ensure that they meet the requirements for use.

[0057] (5) Aliquot the prepared lysate into appropriate containers, seal them and store them under recommended storage conditions, such as in a refrigerator at 4°C, away from direct sunlight and high temperature environment.

[0058] It should be noted that one substance is completely dissolved before adding another substance to ensure that all raw materials are completely dissolved. The storage conditions are low temperature environment, avoiding direct sunlight and high temperature environment. The specific storage conditions can be reasonably adjusted according to actual conditions, and the purpose is to ensure the activity of proteinase K in plant lysate.

[0059] 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).

[0060] The lysis solution for nucleic acid extraction is used in PCR amplification, specifically, including the following steps:

[0061] (1) Take fresh plant tissue (cut into appropriate size, such as 3×4mm 2 ) and placed in a 1.5 mL centrifuge tube or PCR tube.

[0062] It should be noted that the plant tissue samples can be selected from young leaves, fruits, seeds, roots, and stems of the plant, and the selection is reasonable according to the actual situation. The centrifuge tube or PCR tube is sterile.

[0063] In an optional embodiment of the present invention, the plant tissues selected are fresh Arabidopsis leaves, fresh rice leaves and fresh corn leaves, and the growth period corresponds to healthy leaves (light green or bright green without wilting) in the seedling stage or flowering stage of each plant tissue. In other embodiments of the present invention, other parts of the plant tissue can be selected as needed.

[0064] (2) Add 10 μL-200 μL of lysis buffer to the tube and grind the plant tissue thoroughly with a grinding tool until no large pieces are left.

[0065] It should be noted that the grinding tool is a sterile grinding tool.

[0066] (3) Place the tube in a 50-55°C water bath and incubate for 10-30 min, then transfer to a 90-98°C water bath and heat for 5-10 min to inactivate proteinase K.

[0067] It should be noted that the temperature is raised to 90°C-98°C to inactivate the enzyme in the lysate to prevent interference with subsequent PCR amplification.

[0068] (4) The system after water bath treatment is centrifuged and the supernatant is directly used as a PCR amplification template.

[0069] It should be noted that the centrifugation treatment is mainly used to separate the supernatant from the suspended matter in the system, so as to facilitate the separation of the supernatant and direct PCR amplification treatment.

[0070] (5) Preparing a PCR reaction system, including template DNA, PCR reagents, PCR primers, and deionized water.

[0071] 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 Biotechnology (Shanghai) Co., Ltd. (specifically named 2X HyperMB Taq PCR Master Mix Pro), the template DNA is the cleavage product, the PCR primers are shown in Table 1, and the amplification system is shown in Table 2.

[0072] Table 1 PCR primer information

[0073]

[0074] Table 2 Amplification system components

[0075]

[0076]

[0077] (6) Place the PCR reaction system in a PCR instrument and perform amplification according to the preset PCR amplification program. The amplification program is shown in Table 3:

[0078] Table 3 Amplification procedures

[0079]

[0080] (7) After amplification, detect the PCR product by electrophoresis or other appropriate methods.

[0081] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0082] Example 1

[0083] This embodiment provides a lysate for nucleic acid extraction, which includes the following preparation method:

[0084] (1) First, add 65 mL of deionized water into a clean beaker.

[0085] (2) Add the lysis solution components, stirring each substance thoroughly until it is completely dissolved before adding another substance.

[0086] The lysate components include: 50 mM lithium hydroxide (LiOH), 2 M betaine, 0.1 w / v% nonylphenol polyoxyethylene ether, 2 w / v% sodium dodecyl sulfate (SDS), and 10% polysucrose 70.

[0087] (3) Finally, add proteinase K to 0.3 mg / mL and stir thoroughly again.

[0088] (4) Add deionized water to make the final volume 100 ml and check the pH value to 8.0 to ensure that it meets the requirements for use.

[0089] (5) The prepared lysate was dispensed into appropriate containers, sealed and stored in a 4°C refrigerator.

[0090] Experimental Example 1

[0091] This experimental example provides an application of a lysis solution for nucleic acid extraction in direct PCR amplification, as follows:

[0092] (1) Take a fresh Arabidopsis leaf, 3×4 mm in size 2 , placed in a 1.5mL centrifuge tube.

[0093] (2) Add 100 μL of the lysis solution prepared in Example 1 to a centrifuge tube, and grind the leaves thoroughly with a sterile grinding rod until the tissue is completely lysed.

[0094] (3) Place the centrifuge tube in a 50°C water bath and incubate for 30 min. Then transfer to a 95°C water bath and continue heating for 5 min to inactivate proteinase K.

[0095] (4) After a brief centrifugation, take the supernatant directly as a PCR template.

[0096] (5) Prepare a PCR reaction system, including template DNA (Arabidopsis leaf lysate obtained in (4)), 2×PCR mix, PCR primers (At-500bp-F / R) and deionized water. Specific PCR primer information is shown in Table 1, and the amplification system is shown in Table 2.

[0097] (6) Place the PCR reaction system into a PCR instrument and perform amplification according to the preset PCR program. The specific amplification program is shown in Table 3.

[0098] (7) After amplification, the PCR product is detected by electrophoresis.

[0099] Experimental Example 2

[0100] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 in that:

[0101] (1) Take a fresh rice leaf, size 3×4 mm 2 , placed in a 1.5mL centrifuge tube.

[0102] Experimental Example 3

[0103] This experimental example provides an application of a lysis solution for nucleic acid extraction in PCR amplification, which differs from Experimental Example 1 in that:

[0104] (1) Take a fresh corn leaf, size 3×4mm 2 , placed in a 1.5mL centrifuge tube.

[0105] Test Example 1

[0106] This test example studies the final concentration of lithium hydroxide (LiOH) in the lysate for nucleic acid extraction, and the specific implementation process is as follows:

[0107] (1) In a clean beaker, first add deionized water to 65% of the required volume.

[0108] (2) Add the lysis buffer components in sequence, stirring thoroughly after each addition until completely dissolved.

[0109] The lysate components include: concentration gradient lithium hydroxide (LiOH), 2M betaine, 0.1w / v% nonylphenol polyoxyethylene ether, 2w / v% sodium dodecyl sulfate (SDS), and 10% polysucrose 70.

[0110] The concentrations of lithium hydroxide were 10 mM, 30 mM, 50 mM, 70 mM and 100 mM.

[0111] (3) Finally, add proteinase K to 0.3 mg / mL and stir thoroughly again.

[0112] (4) Add deionized water to make the final volume 100 ml and check the pH value to 8.0.

[0113] (5) The prepared lysate was dispensed into appropriate containers, sealed and stored in a 4°C refrigerator.

[0114] Test Example 2

[0115] 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:

[0116] (2) Add the lysis buffer components and stir thoroughly after each addition until completely dissolved.

[0117] The lysate components include: 50 mM lithium hydroxide (LiOH), 2 M betaine, 0.1 w / v% nonylphenol polyoxyethylene ether, concentration gradient sodium dodecyl sulfate (SDS), and 10% polysucrose 70.

[0118] The concentrations of SDS were 1 w / v%, 2 w / v%, 3 w / v%, 4 w / v% and 5 w / v%.

[0119] Test Example 3

[0120] 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:

[0121] (3) Finally, add proteinase K with a concentration gradient and stir thoroughly again.

[0122] Among them, the concentrations of proteinase K are 0.1 mg / mL, 0.3 mg / mL, 0.5 mg / mL and 0.7 mg / mL respectively.

[0123] Test Example 4

[0124] This test example studies the final concentration of betaine in the lysis solution used for nucleic acid extraction. The specific implementation process is different from that of Test Example 1 only in that:

[0125] (2) Add the lysis buffer components and stir thoroughly after each addition until completely dissolved.

[0126] The lysate components include: 50 mM lithium hydroxide (LiOH), concentration gradient betaine, 0.1 w / v% nonylphenol polyoxyethylene ether, 2 w / v% sodium dodecyl sulfate (SDS), and 10% polysucrose 70.

[0127] The concentrations of betaine are 1M, 2M, 3M and 4M respectively.

[0128] Test Example 5

[0129] 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:

[0130] (2) Add the lysis buffer components and stir thoroughly after each addition until completely dissolved.

[0131] The lysate components include: 50 mM lithium hydroxide (LiOH), 2 M betaine, concentration gradient nonylphenol polyoxyethylene ether, 2 w / v% sodium dodecyl sulfate (SDS), and 10% polysucrose 70.

[0132] The concentrations of nonylphenol polyoxyethylene ether are 0.05 w / v%, 0.1 w / v%, 0.15 w / v% and 0.2 w / v% respectively.

[0133] Test Example 6

[0134] 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:

[0135] (2) Add the lysis buffer components and stir thoroughly after each addition until completely dissolved.

[0136] The lysate components include: 50 mM lithium hydroxide (LiOH), 2 M betaine, 0.1 w / v% nonylphenol polyoxyethylene ether, 2 w / v% sodium dodecyl sulfate (SDS), and concentration gradient polysucrose 70.

[0137] The concentrations of polysucrose 70 are 5%, 7%, 10%, 13% and 15% respectively.

[0138] Test Example 7

[0139] This test example detects the PCR amplification products in Experimental Examples 1-3 and Experimental Examples 1-6, and specifically evaluates the performance of the lysate by electrophoresis detection. 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.

[0140] The test results are as follows: Figure 1 This is the result of Arabidopsis leaf lysis amplification; Figure 2 This is the result of rice leaf lysis amplification; Figure 3 This is the result of maize leaf lysis amplification.

[0141] from Figure 1 , Figure 2 and Figure 3 It can be seen that for Arabidopsis leaves and rice leaves, clear and specific PCR amplification products can be observed by electrophoresis detection, indicating that the lysis solution for nucleic acid extraction prepared by the present invention can successfully lyse these two plant tissues and be directly used for PCR amplification.

[0142] Figure 4 The results of screening for LiOH in the lysate; Figure 5 The results of SDS screening in lysate; Figure 6 The results are for the screening of proteinase K in the lysate; Figure 7The results of screening for betaine in the lysate are shown in Figure 2. Figure 8 The results are the screening results of nonylphenol polyoxyethylene ether in the pyrolysis solution; Fig. 9 The results are shown for the screening of polysucrose-70 in the lysate.

[0143] from Figure 4-Figure 9 It can be clearly seen from the results that the core components of the tissue lysate involved in the present invention are lithium hydroxide (or alkaline substances such as potassium hydroxide), SDS, proteinase K, and betaine, and nonylphenol polyoxyethylene ether and polysucrose 70 mainly play a promoting role in enhancing the effect.

[0144] In summary, the lysate for nucleic acid extraction provided by the embodiment of the present invention can successfully lyse Arabidopsis leaves, rice leaves and corn leaves, and can be directly used for PCR amplification to obtain clear and specific PCR amplification products. When in use, only a small amount of plant tissue needs to be added to the lysate, and after simple treatment, PCR amplification can be directly performed without the need for cumbersome DNA extraction and purification steps. It can quickly and efficiently lyse plant cells and release a large amount of high-quality DNA. The lysate has good compatibility with the PCR amplification system and can be directly used for PCR amplification to obtain clear and specific amplification products.

[0145] 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: 10mM-100mM alkaline substance, 0.05w / v%-6w / v% compound surfactant, 0.1mg / mL-0.5mg / mL proteinase K and compound osmotic agent; Wherein, the compound surfactant comprises nonylphenol polyoxyethylene ether and sodium lauryl sulfate; The compound penetrant comprises 1M-4M of penetrant A and 5w / v%-15w / v% of penetrant B.

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 lithium hydroxide.

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

4. The lysate according to claim 1, characterized in that The final concentration of the osmotic agent B is 7 w / v%-13 w / v%, and the osmotic agent B is polysucrose 70.

5. The lysate according to claim 1, characterized in that The final concentration of nonylphenol polyoxyethylene ether in the composite surfactant is 0.05 w / v%-0.2 w / v%, and the final concentration of sodium dodecyl sulfate is 1.5 w / v%-2.5 w / v%.

6. The lysate according to claim 1, characterized in that The final concentration of the proteinase K is 0.2 mg / mL-0.4 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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