CN114350447A - Nucleic acid pollution scavenger and preparation method thereof - Google Patents
Nucleic acid pollution scavenger and preparation method thereof Download PDFInfo
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- CN114350447A CN114350447A CN202210026958.2A CN202210026958A CN114350447A CN 114350447 A CN114350447 A CN 114350447A CN 202210026958 A CN202210026958 A CN 202210026958A CN 114350447 A CN114350447 A CN 114350447A
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Abstract
The invention discloses a nucleic acid pollution scavenger and a preparation method thereof, and relates to the technical field of biology. The nucleic acid pollution scavenger consists of the following raw materials in percentage by mass: 0.4-1.0% of peroxide, 3-4% of ethanol, 0.2-0.4% of isopropanol, 0.2-0.8% of surfactant, 0.1-1.0% of cracking agent, 0.01-0.1% of metal salt, 0.1-1.0% of polyethylene glycol and the balance of deionized water. The nucleic acid pollution scavenger has no corrosive performance, and has the characteristics of obvious scavenging effect and low cost.
Description
Technical Field
The invention particularly relates to a nucleic acid pollution scavenger and a preparation method thereof.
Background
With the development of molecular biology, nucleic acid-centered technologies such as nucleic acid editing, amplification, sequencing, identification, etc. have become more advanced and convenient. Especially, single cell RNA sequencing, digital PCR and transgenic technology are emerging, so that the application of nucleic acid technology is more and more extensive, and the sensitivity is also more and more high. Nucleic acid is a generic term for deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), and is a biological macromolecular compound synthesized by polymerizing many nucleotide monomers, which is one of the most basic substances of life. Nucleic acid detection is a scientific experiment with high accuracy, and pollution is an inevitable problem, wherein pollution caused by the experimental environment is pollution which is the most easily generated and the most difficult to eliminate.
In the daily work of laboratory, laboratory apparatus, instrument, equipment frequency of use is high, and the condition such as the disinfection of appearing not in time or washing and disinfecting thoroughly causes instrument, equipment cross contamination to probably cause the laboratory aerosol pollution when serious, make the experimental result appear the deviation or even mistake, consequently effectively handle nucleic acid pollution reluctantly.
The existing commercial nucleic acid pollution cleaning agent on the market has the defects of high production cost, over-low capacity, high requirement on storage conditions, strong corrosivity and the like, so that the problem to be solved is urgently needed to provide the nucleic acid pollution cleaning agent which is low in cost and corrosivity and is convenient for large-scale mass production.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a nucleic acid pollution scavenger which has low cost, no corrosiveness and good nucleic acid scavenging effect.
The nucleic acid pollution scavenger consists of the following raw materials in percentage by mass:
0.4 to 1.0 percent of peroxide,
3 to 4 percent of ethanol,
0.2 to 0.4 percent of isopropanol,
0.2 to 0.8 percent of surfactant,
0.1 to 1.0 percent of cracking agent,
0.01 to 0.1% of a metal salt,
0.1 to 1.0 percent of polyethylene glycol,
the balance of deionized water.
Preferably, the peroxide is at least one of hydrogen peroxide, peracetic acid, or zinc peroxide.
Preferably, the surfactant is cationic.
Further preferably, the cationic surfactant is a quaternary ammonium salt surfactant.
Preferably, the metal salt is at least one of a copper salt or a zinc salt.
Further preferably, the copper salt is at least one of copper nitrate, copper chloride and copper sulfate, and the zinc salt is at least one of zinc nitrate, zinc sulfate and zinc chloride.
Preferably, the polyethylene glycol is at least one of polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600 and polyethylene glycol 800.
Preferably, the cleavage agent is moroxydine hydrochloride.
Another object of the present invention is to provide a method for preparing the nucleic acid contamination scavenger, comprising the steps of:
(1) weighing raw materials according to a proportion, and then mixing peroxide and deionized water to obtain a first mixture;
(2) adding a surfactant into the first mixture, fully mixing, adding polyethylene glycol, ethanol and isopropanol, and uniformly stirring to obtain a second mixture;
(3) and sequentially adding a cracking agent and metal salt into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Compared with the prior art, the invention has the following beneficial effects:
the nucleic acid pollution scavenger provided by the invention has low peroxide content, so that the corrosion performance of the scavenger is reduced. In order to improve the treatment effect of the scavenging agent on nucleic acid pollution, the invention also adds a certain proportion of metal salt, cracking agent and other components, thereby ensuring the scavenging effect. The nucleic acid pollution scavenger has low cost and is convenient for industrial mass production.
Detailed Description
The invention provides a nucleic acid pollution scavenger which is prepared from the following raw materials in percentage by mass:
0.4-1.0% of peroxide, 3-4% of ethanol, 0.2-0.4% of isopropanol, 0.2-0.8% of surfactant, 0.1-1.0% of cracking agent, 0.01-0.1% of metal salt, 0.1-1.0% of polyethylene glycol and the balance of deionized water.
In some embodiments, the peroxide of the present invention is at least one of hydrogen peroxide, peracetic acid, or zinc peroxide.
In some embodiments, the surfactant in the nucleic acid contamination remover of the present invention is a cationic surfactant.
In some embodiments, the cationic surfactant in the nucleic acid contamination remover of the present invention is a quaternary ammonium salt surfactant.
In some embodiments, the metal salt in the nucleic acid contamination scavenger of the present invention is at least one of a copper salt or a zinc salt.
In some embodiments, the copper salt in the nucleic acid contamination removing agent of the present invention is at least one of copper nitrate, copper chloride and copper sulfate, and the zinc salt is at least one of zinc nitrate, zinc sulfate and zinc chloride.
In some embodiments, the polyethylene glycol in the nucleic acid contamination remover of the present invention is at least one of polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, and polyethylene glycol 800.
In some embodiments, the cleaving agent in the nucleic acid contamination remover of the present invention is moroxydine hydrochloride.
The preparation method of the nucleic acid pollution scavenger comprises the following steps:
(1) weighing raw materials according to a proportion, and then mixing peroxide and deionized water to obtain a first mixture;
(2) adding a surfactant into the first mixture, fully mixing, adding polyethylene glycol, ethanol and isopropanol, and uniformly stirring to obtain a second mixture;
(3) and sequentially adding a cracking agent and metal salt into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
The present invention will be further described with reference to the following specific examples.
Example 1
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.8% of hydrogen peroxide, 3.5% of ethanol, 0.3% of isopropanol, 0.6% of quaternary ammonium salt surfactant, 0.7% of moroxydine hydrochloride, 0.5% of copper nitrate, 0.01% of zinc nitrate, 6000.4% of polyethylene glycol and the balance of deionized water, wherein the hydrogen peroxide and the deionized water are firstly mixed to obtain a first mixture;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 600, ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and (3) adding moroxydine hydrochloride, copper nitrate and zinc nitrate into the mixture II in sequence, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Example 2
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.5% of peroxyacetic acid, 3.5% of ethanol, 0.3% of isopropanol, 0.5% of quaternary ammonium salt surfactant, 0.5% of moroxydine hydrochloride, 0.3% of copper chloride, 0.5% of zinc chloride, 4000.5% of polyethylene glycol and the balance of deionized water, and firstly, mixing the peroxyacetic acid with the deionized water to obtain a mixture I;
(2) adding a surfactant into the first mixture, fully mixing, adding polyethylene glycol 400, ethanol and isopropanol, and uniformly stirring to obtain a second mixture;
(3) and (3) adding moroxydine hydrochloride, copper chloride and zinc chloride into the mixture II in sequence, and stirring uniformly to obtain the nucleic acid pollution scavenger.
Example 3
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
1.0% of zinc peroxide, 4% of ethanol, 0.2% of isopropanol, 0.8% of quaternary ammonium salt surfactant, 1.0% of moroxydine hydrochloride, 0.5% of copper sulfate, 0.5% of zinc chloride, 2000.1% of polyethylene glycol and the balance of deionized water, and firstly mixing the zinc peroxide and the deionized water to obtain a mixture I;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 200, ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and (3) adding moroxydine hydrochloride, copper sulfate and zinc chloride into the mixture II in sequence, and stirring uniformly to obtain the nucleic acid pollution scavenger.
Example 4
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
1.0% of hydrogen peroxide, 4% of ethanol, 0.3% of isopropanol, 0.6% of quaternary ammonium salt surfactant, 0.7% of moroxydine hydrochloride, 0.5% of copper nitrate, 0.01% of zinc nitrate, 6000.4% of polyethylene glycol and the balance of deionized water, and firstly mixing the hydrogen peroxide and the deionized water to obtain a mixture I;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 600, ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and (3) adding moroxydine hydrochloride, copper nitrate and zinc nitrate into the mixture II in sequence, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Example 5
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.4% of hydrogen peroxide, 3% of ethanol, 0.2% of isopropanol, 0.3% of quaternary ammonium salt surfactant, 0.1% of moroxydine hydrochloride, 0.5% of copper chloride, 6000.4% of polyethylene glycol and the balance of deionized water, wherein the hydrogen peroxide and the deionized water are firstly mixed to obtain a first mixture;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 600, ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and sequentially adding moroxydine hydrochloride and copper chloride into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Example 6
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.7% of hydrogen peroxide, 4% of ethanol, 0.3% of isopropanol, 0.6% of quaternary ammonium salt surfactant, 0.7% of moroxydine hydrochloride, 1.0% of zinc nitrate, 8000.4% of polyethylene glycol and the balance of deionized water, wherein the hydrogen peroxide and the deionized water are firstly mixed to obtain a first mixture;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 800, ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and sequentially adding moroxydine hydrochloride and zinc nitrate into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Comparative example 1
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.8% of hydrogen peroxide, 3.5% of ethanol, 0.6% of quaternary ammonium salt surfactant, 0.7% of moroxydine hydrochloride, 0.5% of copper nitrate, 6000.4% of polyethylene glycol and the balance of deionized water, and firstly mixing the hydrogen peroxide and the deionized water to obtain a first mixture;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding polyethylene glycol 600 and ethanol, and uniformly stirring to obtain a mixture II;
(3) and sequentially adding moroxydine hydrochloride into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
Comparative example 2
A nucleic acid pollution scavenger is prepared by the following steps:
(1) weighing the following raw materials in percentage by weight:
0.8% of hydrogen peroxide, 3.5% of ethanol, 0.3% of isopropanol, 0.6% of quaternary ammonium salt surfactant, 0.7% of moroxydine hydrochloride and the balance of deionized water, wherein the hydrogen peroxide and the deionized water are firstly mixed to obtain a mixture I;
(2) adding a quaternary ammonium salt surfactant into the mixture I, fully mixing, adding ethanol and isopropanol, and uniformly stirring to obtain a mixture II;
(3) and sequentially adding moroxydine hydrochloride into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
The scavenger prepared by the above examples and comparative examples was tested as follows:
the reagents described in table 1 were sequentially loaded into a centrifuge tube, then placed in an ultrasonic instrument for ultrasonic treatment for 30min, mixed uniformly, centrifuged instantly, and left to stand for 5 min.
And finally, detecting the sample in the centrifugal tube by using 1% agarose gel electrophoresis, and observing under an ultraviolet lamp.
TABLE 1
| Reagent | Amount of the composition used |
| TBGreenPremixExTaqⅡ(2X) | 12.5μL |
| PCRForwardPrimer(20μM) | 0.5μL |
| PCRReversePrimer(20μM) | 0.5μL |
| Lambda DNA template (1. mu.g/mL) | 2.5μL |
| Sterilized water | 7.2μL |
| Nucleic acid pollution scavenger | 1.8μL |
It was found by experiment that the samples containing the scavengers described in examples 1-6 had no bright band under the UV lamp, while the samples containing the scavengers described in comparative examples 1-2 had a different degree of bright band under the UV lamp. It is shown that the scavengers prepared in comparative examples 1-2 have a poor effect of eliminating nucleic acid contamination. The scavenging agent prepared in the examples 1-6 of the invention has far higher effect on scavenging nucleic acid pollution than the comparative example.
It should be noted that the above-mentioned embodiments are merely examples of the present invention, and it is obvious that the present invention is not limited to the above-mentioned embodiments, and other modifications are possible. All modifications directly or indirectly derivable by a person skilled in the art from the present disclosure are to be considered within the scope of the present invention.
Claims (9)
1. The nucleic acid pollution scavenger is characterized by comprising the following raw materials in percentage by mass:
0.4 to 1.0 percent of peroxide,
3 to 4 percent of ethanol,
0.2 to 0.4 percent of isopropanol,
0.2 to 0.8 percent of surfactant,
0.1 to 1.0 percent of cracking agent,
0.01 to 0.1% of a metal salt,
0.1 to 1.0 percent of polyethylene glycol,
the balance of deionized water.
2. The nucleic acid contamination eliminating agent according to claim 1, wherein the peroxide is at least one of hydrogen peroxide, peracetic acid, or zinc peroxide.
3. The nucleic acid contamination removing agent according to claim 1, wherein the surfactant is a cationic surfactant.
4. The nucleic acid contamination removing agent according to claim 3, wherein the cationic surfactant is a quaternary ammonium salt-based surfactant.
5. The nucleic acid contamination scavenger according to claim 1, wherein the metal salt is at least one of a copper salt or a zinc salt.
6. The nucleic acid contamination scavenger according to claim 5, wherein the copper salt is at least one of copper nitrate, copper chloride and copper sulfate, and the zinc salt is at least one of zinc nitrate, zinc sulfate and zinc chloride.
7. The nucleic acid contamination removing agent according to claim 1, wherein the polyethylene glycol is at least one of polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, and polyethylene glycol 800.
8. The nucleic acid contamination scavenger according to claim 1, wherein the cleaving agent is moroxydine hydrochloride.
9. The method for producing a nucleic acid contamination removing agent according to any one of claims 1 to 8, comprising the steps of:
(1) weighing raw materials according to a proportion, and then mixing peroxide and deionized water to obtain a first mixture;
(2) adding a surfactant into the first mixture, fully mixing, adding polyethylene glycol, ethanol and isopropanol, and uniformly stirring to obtain a second mixture;
(3) and sequentially adding a cracking agent and metal salt into the mixture II, and uniformly stirring to obtain the nucleic acid pollution scavenger.
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| CN115786045A (en) * | 2022-12-02 | 2023-03-14 | 量准（上海）医疗器械有限公司 | Laboratory nucleic acid pollution scavenger and application thereof |
| CN115786039A (en) * | 2023-02-13 | 2023-03-14 | 北京迈佳致和科技有限公司 | Nucleic acid scavenger based on silver and preparation method and application thereof |
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| CN115786039A (en) * | 2023-02-13 | 2023-03-14 | 北京迈佳致和科技有限公司 | Nucleic acid scavenger based on silver and preparation method and application thereof |
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