A nucleic acid scavenger, its preparation and use

Abstract

The present application provides a nucleic acid scavenger and its preparation method and application. The nucleic acid scavenger comprises an organic acid, a metal salt ion, a peroxide, a nucleic acid precipitant and a surfactant. The present application can quickly and effectively remove nucleic acid pollution in equipment consumables and laboratory environment, avoiding false positive of experimental results. At the same time, the components used are non-toxic and harmless, and will not cause harm to the experimental personnel when used, and will not cause pollution to the overall environment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of nucleic acid detection / amplification, and particularly relates to a nucleic acid scavenger as well as a preparation method and application thereof, which can be used in a matched PCR technology and related application scenarios. BACKGROUND

[0002] PCR, full name Polymerase chain reaction, is a molecular biology technology that can amplify specific DNA fragments in vitro through DNA double-strand replication principle by DNA polymerase and primers, which can enrich a large number of copies from a trace amount of DNA template. It provides a favorable basis for qualitative and quantitative analysis of experimental objects. Therefore, PCR technology is widely used in many fields of biology and related disciplines, and is more applied in medical research, crime evidence and molecular archaeology.

[0003] Because PCR has high detection sensitivity and high enrichment in time, its main disadvantage is easy to be contaminated. Nucleic acid laboratory contamination mainly includes the following types: sample contamination, reagent contamination, environmental contamination, equipment contamination and other contaminations. Sample and reagent contamination is relatively simple to exclude, and the sample and reagent can be replaced or sent to other laboratories for verification, and environmental contamination and equipment contamination need to be thoroughly disinfected and cleaned.

[0004] In order to ensure the safety of the experimenters and save the valuable time of the experimenters, it is necessary to invent a non-toxic and harmless, time-saving and labor-saving reagent to help quickly and effectively remove nucleic acid contamination in the laboratory and improve the accuracy of the experiment. SUMMARY

[0005] In order to solve the problem of lacking non-toxic and harmless, time-saving and labor-saving reagents to quickly and effectively remove nucleic acid contamination, the present application provides a high-efficiency nucleic acid scavenger as well as a preparation method and application thereof, which is non-toxic and harmless to personnel and environment, can be used for instruments and equipment, is convenient to operate, and has a rapid and significant effect.

[0006] To solve the above technical problems, the present application provides a nucleic acid scavenger in the first aspect, which comprises: an organic acid, a metal salt ion, a peroxide, a nucleic acid precipitant and a surfactant; the concentration of the peroxide is greater than or equal to 1.5%.

[0007] In some embodiments, the organic acid is selected from one or more of erythorbic acid, ascorbic acid, malic acid, citric acid, ethylenediaminetetraacetic acid and succinic acid; preferably, the organic acid is erythorbic acid, malic acid and / or citric acid.

[0008] In some embodiments, the nucleic acid scavenger satisfies one or more of the following conditions:

[0009] said metal salt ion is selected from one or more of Cu 2+ , Fe 3+ , Fe 2+ , Co 2+ ;

[0010] said nucleic acid precipitating agent is polyethylene glycol or polyethylene imine;

[0011] said surfactant is octoxynol;

[0012] said peroxide is selected from one or more of hydrogen peroxide, sodium peroxide, and peroxoacetic acid.

[0013] Preferably, said metal salt ion is Cu 2+ , Fe 2+ , and / or Co 2+ ; and said peroxide is hydrogen peroxide and / or sodium peroxide.

[0014] In certain embodiments, said organic acid is present in an amount greater than or equal to 0.01 mol / L, preferably from 0.005 to 0.15 mol / L; more preferably from 0.01 to 0.15 mol / L; and even more preferably from 0.01 to 0.03 mol / L.

[0015] In certain embodiments, said metal salt ion is present in an amount greater than or equal to 0.005 mol / L, preferably from 0.0025 to 0.15; more preferably from 0.005 to 0.15 mol / L; and even more preferably from 0.005 to 0.0125 mol / L.

[0016] In certain embodiments, said nucleic acid precipitating agent is present in an amount greater than or equal to 0.5 g / L, preferably from 0.1 to 3.0 g / L, and more preferably from 0.5 to 2.0 g / L.

[0017] In certain embodiments, said surfactant is present in an amount greater than or equal to 0.01 %, preferably from 0.01 % to 0.1 %, and more preferably from 0.01 % to 0.04 %.

[0018] In certain embodiments, when said peroxide is sodium peroxide, said peroxide is present in an amount greater than or equal to 2.5 %, preferably from 2.5 % to 5 %, and more preferably from 2.5 % to 3.5 %.

[0019] When said peroxide is hydrogen peroxide, said peroxide is present in an amount greater than or equal to 1.5 %, preferably from 1.5 % to 5 %, and more preferably from 1.5 % to 3 %.

[0020] The second aspect of the present application provides a kit comprising the nucleic acid scavenger according to the first aspect of the present application. Preferably, the kit further comprises at least one of a positive control sample, Taq enzyme, up and down stream primers for a housekeeping gene of human genome and / or buffer for amplification.

[0021] The third aspect of the present application provides a method for scavenging nucleic acid contamination, which comprises mixing or contacting a sample with the nucleic acid scavenger according to the first aspect of the present application.

[0022] Preferably, the sample is a human genome sample, a plasmid sample, a lambda DNA sample, MS 2, an RNA sample and / or an amplification product sample containing nucleic acid.

[0023] The fourth aspect of the present application provides use of the nucleic acid scavenger according to the first aspect of the present application in scavenging nucleic acid or preparing a kit for scavenging nucleic acid.

[0024] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined in any manner to obtain various preferred embodiments of the present application.

[0025] The reagents and raw materials used in the present application are commercially available.

[0026] The positive progress effect of the present application is that:

[0027] The nucleic acid scavenger provided by the present application has the characteristics of high efficiency, rapidity and safety. It can completely remove DNA, RNA, PCR amplification product and inactivation of bacteriophage, bacteria, fungi and viruses; the components are degradable and non-corrosive, and can be directly sprayed on the surface of instruments and equipment without harm to humans. The contamination can be easily removed by spraying liquid for 2-10 minutes. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 The figure is the removal effect of Puc19 plasmid sample by embodiments 1-3.

[0029] Figure 2 The figure is the removal effect of lambda DNA sample by embodiments 1-3.

[0030] Figure 3 The figure is the removal effect of human genome sample by embodiments 1-3.

[0031] Figure 4 The figure is the removal effect of MS 2 RNA sample by embodiments 1-3.

[0032] Figure 5 The figure is the removal effect of 100 bp amplification product sample by embodiments 1-3.

[0033] Figure 6Figure 1. Results of placing amplification system with open lid on clean bench without any treatment of amplification product.

[0034] Figure 7 Figure 2. Results of placing amplification system with open lid on clean bench after treatment of amplification product according to embodiments 1-3.

[0035] Figure 8 Figure 3. Results of recovering template from human genome coated on lab bench without treatment and after treatment according to embodiments.

[0036] Figure 9 Figure 4. Removal of 100 bp amplification product by embodiment 1 at different peroxide concentrations.

[0037] Figure 10 Figure 5. Removal of 100 bp amplification product by embodiment 3 at different peroxide concentrations.

[0038] Figure 11 Figure 6. Removal of 100 bp amplification product by embodiment 2 at different peroxide concentrations.

[0039] Figure 12 Figure 7. Removal of 100 bp amplification product by embodiment 2 at different metal ion concentrations.

[0040] Figure 13 Figure 8. Removal of 100 bp amplification product by embodiment 2 formulated with different organic acids.

[0041] Figure 14 Figure 9. Removal of 100 bp amplification product by embodiment 2 at different concentrations of organic acids.

[0042] Figure 15 Figure 10. Removal of 100 bp amplification product by embodiment 2 at different concentrations of nucleic acid precipitants or different concentrations of surfactants.

[0043] Figure 16 Figure 11. Removal of Puc19 plasmid samples using embodiments 1-3 after six months of storage.

[0044] Figure 17 Figure 12. Removal of 100 bp amplification product samples using embodiments 1-3 after six months of storage.

[0045] Figure 18 Figure 13. Removal of common nucleic acid contaminants using embodiment 3 after one year of storage. DETAILED DESCRIPTION

[0046] The present application is further illustrated by the following examples without limiting the present application to the described examples. The experimental methods in the following examples, if not otherwise specified, are carried out according to the conventional methods and conditions, or according to the instructions of the commercial products.

[0047] The term "positive", "positive control" or "positive sample" refers to an environment, experimental group or sample contaminated with nucleic acid.

[0048] The term "negative" refers to an environment not contaminated with nucleic acid.

[0049] The mix used in the present application is TaqMan Master Mix with the product number TC001-A from Jiangsu Tongke Biotechnology Co., Ltd. The primers and probes are synthesized by Goldengene Biotechnology Co., Ltd.

[0050] Table 1 Formulation of different nucleic acid scavengers

[0051]

[0052] Example 1 Verification of the removal effect of common sources of contamination in the laboratory

[0053] Common nucleic acid contamination in the laboratory includes: positive control samples of experimental personnel, aerosol leakage of amplification products during the experimental process, etc.

[0054] Experimental design for the above-mentioned situations: verification of the removal of 500 ng human genome samples, 500 ng Puc19 plasmid samples, 500 ng λ DNA samples, 500 ng MS 2 RNA samples, 50 cycles of 100 bp amplification product samples by the implementation scheme.

[0055] Specific operation: In the eight-tube group, four groups are the same sample, that is, take four equal amounts of 500 ng human genome samples, 500 ng Puc19 plasmid samples, 500 ng λ DNA samples, 500 ng MS 2 RNA samples, and 50 cycles of 100 bp amplification product samples, respectively, and place them in the eight-tube group. One part is supplemented with 5 μl of water, and the remaining three parts are added with 5 μl of implementation schemes 1-3. The nucleic acid agarose concentration of the amplification product is 2%, and the nucleic acid agarose concentration of the remaining is 1%. The voltage is 120V and the electrophoresis time is 30 min. During the experiment, the No. 1 hole is the maker, and the No. 2 hole is the untreated control sample. The No. 3-5 holes are the effect pictures after the removal by the implementation schemes 1-3.

[0056] The results are as follows Figures 1-5 In all experiments, except for the marker and the control in the No. 2 hole, there are bands, and the remaining holes have no bands, indicating that the implementation schemes 1-3 can remove common nucleic acid contamination in experiments.

[0057] Example 2 Verification of the cleaning of the environment and equipment during the experiment.

[0058] The frequent opening of the cover of the aerosol and positive control samples during the experiment process leads to the easy pollution of the experimental environment. The amplification product is overflowed by opening the cover and flicking in the clean bench. The prepared amplification system (without template) is placed in the clean bench environment for 3-5 min. The experiment is sealed and performed. After verifying that the product is overflowed by flicking, the clean bench environment is treated with embodiments 1-3 respectively. The prepared amplification system (without template) is placed in the clean bench environment for 3-5 min. The experiment is sealed and performed. The amplification conditions are compared.

[0059] The mix used is TaqMan Master Mix from Jiangsu Tongke Biotechnology Co., Ltd. with the product code TC001-A. The primers and probes are synthesized by Goldengene.

[0060] Divide into four groups, each group with 8 repeats. First prepare the amplification system in the negative interval: add 12.5 μl MIX (containing Taq enzyme), 1 μl 10 μM upstream primer, 1 μl 10 μM downstream primer, 1 μl 10 μM probe, and 9.5 μl water in the eight-tube. One group tests the untreated environment, and the remaining three groups test the environment treated with embodiments 1-3 respectively.

[0061] Prepare the prepared samples for amplification experiment.

[0062] The amplification program is: 95℃ for 10 min, 95℃ for 15 s, 60℃ for 1 min, 45 cycles.

[0063] The results are as follows Figures 6-7 The eight repeats without treatment only one has no obvious amplification and the rest have some amplification. However, after treatment with embodiments 1-3, there is no amplification. It shows that in the contaminated environment, the use of embodiments can achieve good nucleic acid pollution removal effect.

[0064] For the positive samples during the experiment, take four groups of 2 μl 200 ng / μl human genome and spread them on the experimental bench surface of the same material. One group is not treated as a control, and the remaining three groups are treated with embodiments 1-3. Gently wipe the template-coated bench surface with a cotton swab dipped in TE buffer. Shake the cotton swab in 200 μl TE buffer for 2 min to obtain three experimental samples and one control sample. Take 2 μl of the sample as a template for PCR experiment. Compare the amplification conditions before and after treatment.

[0065] Divided into four groups, each group of 3 repeats. First in the negative interval prepared good amplification system: eight in the tube to add 12.5 μl MIX (contains Taq enzyme), 1 μl 10 μM upstream primer, 1 μl 10 μM downstream primer, 1 μl 10 μM probe, 7.5 μl water. One group added 2 μl without treatment recovery template, the remaining three groups were added 2 μl after treatment of the recovery template of scheme 1-3. Prepare to conduct experiments.

[0066] Amplification procedure: 95℃ 10min, 95℃ 15s, 60℃ 1min, 45 cycles.

[0067] The results are shown in Figure 8 The template without treatment after recovery amplification is normal, but the recovery template after treatment of scheme 1-3 has no amplification, which shows that scheme 1-3 has good effect on the removal of nucleic acid residues of experimental equipment.

[0068] Example 3 removal effect of different peroxide concentrations

[0069] Change the peroxide concentration of experimental scheme 1-3, and verify the removal effect of scheme 1-3 under different peroxide concentrations on 50 cycles of 100 bp amplification product.

[0070] The specific operation is: set the peroxide concentration of scheme 1 and scheme 3 to 1.0%, 1.5%, 2%, 2.5%, 3% and 3.5% gradient concentration respectively, and the rest of the formula is consistent with scheme 1 and scheme 3 of table 1. Set the peroxide concentration of scheme 2 to 0.25%, 0.5%, 1%, 1.5%, 2% and 3% gradient. The rest of the formula is consistent with scheme 2 of table 1.

[0071] Add equal amount of 50 cycles of 100 bp amplification product sample twenty-six in eight tubes, three add 5 μl water as untreated control, seven add 5 μl of different concentration peroxide scheme 1, seven add 5 μl of different concentration peroxide scheme 3. The concentration of nucleic acid gel agarose is 3%, and the voltage is 120V electrophoresis for 30min. The experiment is divided into three pieces of gel:

[0072] Gel 1 with No. 1 hole as maker, No. 2-3 hole as untreated control sample. No. 4-5 hole for scheme 1 and scheme 3 removal effect of peroxide concentration of 1%; No. 6-7 hole for scheme 1 and scheme 3 removal effect of peroxide concentration of 1.5%; No. 8 and No. 9 hole for scheme 1 and scheme 3 removal effect of peroxide concentration of 2%.

[0073] Gel 2 with No. 1 hole as maker, No. 2-3 hole as untreated control sample. No. 4-5 hole as the removal effect of embodiment 1 and embodiment 3 with peroxide concentration of 2.5%; No. 6-7 hole as the removal effect of embodiment 1 and embodiment 3 with peroxide concentration of 3%; No. 8 and No. 9 hole as the removal effect of embodiment 1 and embodiment 3 with peroxide concentration of 3.5%.

[0074] Gel 3 with No. 1 hole as maker, No. 2 hole as untreated control sample; No. 3 hole as the removal effect of embodiment 2 with peroxide concentration of 0.25%; No. 4 hole as the removal effect of embodiment 2 with peroxide concentration of 0.5%; No. 5 hole as the removal effect of embodiment 2 with peroxide concentration of 1%; No. 6 hole as the removal effect of embodiment 2 with peroxide concentration of 1.5%; No. 7 hole as the removal effect of embodiment 2 with peroxide concentration of 2%; No. 8 hole as the removal effect of embodiment 2 with peroxide concentration of 3%.

[0075] The results are shown in Table 4. Figures 9-11 As shown in Table 4, the effect of embodiment 1 and embodiment 3 on the treatment of amplification product showed an increasing trend at peroxide concentration of 1%, 1.5%, and 2%, but could not completely remove the amplification product, and could completely remove the amplification product at peroxide concentration of 2.5% and above. The effect of experimental scheme 2 on the treatment of amplification product showed an increasing trend at peroxide concentration of 0.25% and 0.5%, but could not completely remove the amplification product, and could completely remove the amplification product at peroxide concentration of 1% and above.

[0076] Example 4 Removal effect of different metal ion concentrations

[0077] The metal ion concentration of experimental scheme 2 was changed to verify the removal effect of embodiment 2 on 50 cycles of 100 bp amplification product at different metal ion concentrations.

[0078] The specific operation is as follows: the metal ion concentration in embodiment 2 is set to gradient concentrations of 0.0025 mol / L, 0.005 mol / L, 0.0075 mol / L, 0.01 mol / L, and 0.0125 mol / L, and the rest of the formula is consistent with embodiment 2 in Table 1.

[0079] Seven equal volumes of 100bp amplified product samples (50 cycles each) were added to eight tubes. One tube was supplemented with 5 μl of water as an untreated control, and the other five tubes were supplemented with 5 μl of metal ions at different concentrations according to Implementation Plan 2. The nucleic acid gel agarose concentration was 3%, and electrophoresis was performed at 120V for 30 min. Well 1 was used as the marker, well 2 as the untreated control sample, well 3 as the removal effect of Implementation Plan 2 with a metal ion concentration of 0.0025 mol / L, well 4 as the removal effect of Implementation Plan 2 with a metal ion concentration of 0.005 mol / L, well 5 as the removal effect of Implementation Plan 2 with a metal ion concentration of 0.0075 mol / L, well 6 as the removal effect of Implementation Plan 2 with a metal ion concentration of 0.01 mol / L, and well 7 as the removal effect of Implementation Plan 2 with a metal ion concentration of 0.0125 mol / L.

[0080] The results are as follows Figure 12 As shown, the amplification treatment was effective at a metal ion concentration of 0.0025 mol / L, but not completely eliminated the amplification products. At metal ion concentrations of 0.005 mol / L and above, the amplification products were completely eliminated.

[0081] Example 5: Scavenging effect of different organic acids

[0082] In Implementation Scheme 2, malic acid was replaced with different organic acids to verify the scavenging effect of Implementation Scheme 2 on 100bp amplification products in 50 cycles under different organic acid conditions.

[0083] The specific operation is as follows: replace the malic acid in Implementation Scheme 2 with citric acid, succinic acid, ethylenediaminetetraacetic acid, isoascorbic acid, and ascorbic acid of the same concentration for the experiment. The remaining formulas are the same as Implementation Scheme 2 in Table 1.

[0084] Seven equal volumes of 50 cycles of 100bp amplification product samples were added to eight tubes. One tube was supplemented with 5 μl of water as an untreated control, one tube contained 5 μl of the treatment according to Implementation Plan 2, and the remaining five tubes contained 5 μl of different organic acids from Implementation Plan 2. The nucleic acid gel agarose concentration was 2%, and electrophoresis was performed at 120V for 30 min. During the experiment, well 1 was the marker, well 2 contained the untreated 50 cycles of 100bp amplification product sample, and well 3 contained the amplification product sample treated according to Implementation Plan 2. Wells 3-8 contained amplification product samples treated according to Implementation Plan 2 prepared with citric acid, succinic acid, ethylenediaminetetraacetic acid, isoascorbic acid, and ascorbic acid, respectively.

[0085] The results are as follows Figure 13 As shown, different organic acid formulations in Implementation Scheme 2 can effectively remove 100bp amplification product samples over 50 cycles.

[0086] Example 6 Removal effect of different concentrations of organic acid

[0087] The concentration of malic acid in the experimental scheme 2 of Table 1 was changed to verify the removal effect of the implementation scheme 2 at different concentrations on the 50 cycles of 100 bp amplification product.

[0088] Specific operation: set the concentration of organic acid in implementation scheme 2 to 0.005 mol / L, 0.01 mol / L, 0.015 mol / L, 0.02 mol / L, 0.025 mol / L and 0.03 mol / L gradient concentration, and the rest of the formula is consistent with the implementation scheme 2 of Table 1.

[0089] In eight joint tubes, add an equal amount of 50 cycles of 100 bp amplification product sample 7, 1 supplement 5 μl water as untreated control, five 5 μl of different concentration of organic acid implementation scheme 2, nucleic acid gel agarose concentration is 3%, voltage 120V electrophoresis 30 min. With No. 1 hole as maker, No. 2 hole as untreated control sample; No. 3 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.005 mol / L; No. 4 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.01 mol / L; No. 5 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.015 mol / L; No. 6 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.02 mol / L; No. 7 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.025 mol / L; No. 8 hole as the removal effect of implementation scheme 2 with organic acid concentration of 0.03 mol / L.

[0090] The results are shown in Table 2. Figure 14 As shown in Table 2, when the concentration of organic acid is 0.005 mol / L, it has effect on amplification treatment, but it is not completely removed. When the concentration of organic acid is 0.01 mol / L and above, the amplification product can be completely removed.

[0091] Example 7 Removal effect of different concentrations of nucleic acid precipitant and surfactant

[0092] The concentration of nucleic acid precipitant or surfactant in the experimental scheme 2 was changed to verify the removal effect of the implementation scheme 2 on the 50 cycles of 100 bp amplification product at different concentrations of nucleic acid precipitant or surfactant.

[0093] Specific operation: set the concentration of nucleic acid precipitant in implementation scheme 2 to 0.5 g / L, 1 g / L, 1.5 g / L and 2 g / L gradient concentration, and the rest of the formula is consistent with the implementation scheme 2 of Table 1. Set the concentration of surfactant in implementation scheme 2 to 0.01%, 0.02%, 0.03% and 0.04% gradient concentration, and the rest of the formula is consistent with the implementation scheme 2 of Table 1.

[0094] In eight tubes, add equal amounts of 50 cycles of 100 bp amplification product sample 9, 1 supplemented with 5 μl water as untreated control, four of the implementation of 5 μl of different concentrations of nucleic acid precipitant scheme 2, four of the implementation of different concentrations of surfactant scheme 2, nucleic acid gel agarose concentration is 3%, voltage 120V electrophoresis 30 min. With No. 1 hole as maker, No. 2 hole as untreated control sample; No. 3 hole as the implementation of 2 nucleic acid precipitant concentration of 0.5 g / L removal effect; No. 4 hole as the implementation of 2 nucleic acid precipitant concentration of 1 g / L removal effect; No. 5 hole as the implementation of 2 nucleic acid precipitant concentration of 1.5 g / L removal effect; No. 6 hole as the implementation of 2 nucleic acid precipitant concentration of 2 g / L removal effect; No. 7 hole as the implementation of 2 surfactant concentration of 0.01% removal effect; No. 8 hole as the implementation of 2 surfactant concentration of 0.02% removal effect; No. 9 hole as the implementation of 2 surfactant concentration of 0.03% removal effect; No. 10 hole as the implementation of 2 surfactant concentration of 0.04% removal effect.

[0095] The results are shown in Figure 15 The nucleic acid precipitant and surfactant concentration have no effect on the amplification product contamination, and can completely remove the amplification product.

[0096] Example 8 tests the long-term stability of the implementation of 1-3.

[0097] After the implementation of 1-3 is prepared, 500 ng Puc19 plasmid and 50 cycles of 100 bp amplification product are verified for removal after six months.

[0098] The specific operation is: in eight tubes, four groups are the same sample, that is, take 4 equal amounts of 500 ng Puc19 plasmid sample, 50 cycles of 100 bp amplification product sample, respectively, place them in eight tubes, one supplemented with 5 μl water as untreated control, the remaining three added with 5 μl of the implementation of 1-3. The nucleic acid gel agarose concentration of the amplification product is 2%, and the nucleic acid gel agarose concentration of the plasmid is 1%. Voltage 120V electrophoresis 30 min. When the experiment is performed, No. 1 hole is used as maker, No. 2 hole is used as untreated control sample. No. 3-5 hole is the effect diagram after the implementation of 1-3 removal.

[0099] The results are shown in Figures 16-17 The implementation of 1 and the implementation of 3 have some template residues in amplification product treatment, and the implementation of 2 can still effectively remove the two kinds of nucleic acid contamination, which shows that the implementation of 2 still has good effect on nucleic acid contamination removal after six months of preservation.

[0100] After 6 months of incubation, the removal of common nucleic acid contaminants in the laboratory was continued.

[0101] The specific operation is as follows: two groups in the eight-tube are the same sample, that is, two equal amounts of 500 ng human genomic sample, 500 ng Puc19 plasmid sample, 500 ng λ DNA sample, 500 ng MS2 RNA sample, and 50 cycles of 100 bp amplification product sample are respectively placed in the eight-tube, one is supplemented with 5 μl of water as an untreated control, and the other is added with 5 μl of embodiment 2. The concentration of nucleic acid agarose gel is 2% with a voltage of 120 V for electrophoresis for 30 min. During the experiment, the No. 1 hole position is the maker, the No. 2 hole position is the untreated Puc19 sample, the No. 3 hole position is the Puc19 sample treated by embodiment 2, the No. 4 hole position is the λ DNA sample, the No. 5 hole position is the λ DNA sample treated by embodiment 2,

[0102] The No. 6 hole position is the human genomic sample, the No. 7 hole position is the human genomic sample treated by embodiment 2, the No. 8 hole position is the MS2 RNA sample, the No. 9 hole position is the MS2 RNA sample treated by embodiment 2, the No. 10 hole position is the 100 bp amplification product sample, and the No. 11 hole position is the 100 bp amplification product sample treated by embodiment 2.

[0103] The effect diagram after removal is shown in Figure 18 After one year of storage, embodiment 2 can still efficiently remove the five common nucleic acid contaminants.

[0104] The foregoing has described the specific embodiments of the present application in detail, and those of ordinary skill in the art can make various changes, modifications, replacements and variations to these embodiments without departing from the principles and spirits of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A nucleic acid scavenger, characterized in that, The nucleic acid scavenger comprises: an organic acid, a metal salt ion, a peroxide, a nucleic acid precipitant and a surfactant; the peroxide is hydrogen peroxide and / or sodium peroxide; When the peroxide is sodium peroxide, the content of the peroxide is 2.5% to 3.5%; When the peroxide is hydrogen peroxide, the content of the peroxide is 1.5% to 3%; The content of the metal salt ion is 0.005 to 0.0125 mol / L; The content of the organic acid is 0.01 to 0.03 mol / L; The nucleic acid precipitant is polyethylene glycol or polyethylene imine; The surfactant is octylphenoxypolyethoxyethanol.

2. The nucleic acid scavenger of claim 1, wherein The organic acid is selected from one or more of erythorbic acid, ascorbic acid, malic acid, citric acid, ethylenediaminetetraacetic acid and succinic acid.

3. The nucleic acid scavenger of claim 2, wherein The organic acid is erythorbic acid, malic acid and / or citric acid.

4. The nucleic acid scavenger of claim 1, wherein, The metal salt ions are selected from one or more of Cu 2+ , Fe 3+ , Fe 2+ , Co 2+ .

5. The nucleic acid scavenger of claim 4, wherein The metal salt ion is Cu 2+ , Fe 2+ and / or Co 2 + .

6. The nucleic acid scavenger of claim 1, wherein The content of the nucleic acid precipitant is 0.5 to 2.0 g / L.

7. The nucleic acid scavenger of claim 1, wherein The percentage of the surfactant is 0.01% to 0.04%.

8. A kit characterized in that, The kit comprises the nucleic acid scavenger of any one of claims 1 to 7.

9. The kit of claim 8, wherein The kit further comprises at least one of a positive control sample, Taq enzyme, upstream and downstream primers for a housekeeping gene of the human genome and / or a buffer for amplification.

10. A method of removing nucleic acid contamination, characterized by, The method mixes or contacts the sample with the nucleic acid scavenger of any one of claims 1 to 7.

11. The method of claim 10, wherein, The sample is a human genome sample, a plasmid sample, a lambda DNA sample, MS 2, an RNA sample and / or an amplification product sample containing nucleic acid.

12. Use of the nucleic acid scavenger of any one of claims 1 to 7 in scavenging nucleic acid or in preparing a kit for scavenging nucleic acid.

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