Nucleic acid dye and application thereof in nucleic acid detection

Abstract

The invention discloses a nucleic acid dye and application thereof in nucleic acid detection, and belongs to the technical field of nanometer materials and biology, the nucleic acid dye is licorice root derived carbon dots, the carbon dots are prepared based on licorice roots or licorice root components as precursors for the first time, and the carbon dots are successfully applied to the field of nucleic acid detection. The licorice root derived carbon dots replace traditional dyes to be used in agarose gel electrophoresis, and efficient visual detection of DNA, RNA and plasmids can be achieved.

Description

Technical Field

[0001] The invention belongs to the field of nanomaterials and biological technology, and in particular relates to a nucleic acid dye and an application thereof in nucleic acid detection. Background Art

[0002] With the continuous deepening of biological and biochemical research, the study of nucleic acid (DNA and RNA) structure has become particularly important. Nucleic acid dyes, as a compound that can bind to nucleic acids, have become a key tool for observing and analyzing nucleic acids in biological samples. They can help reveal the structure and function of nucleic acids and are an indispensable part of nucleic acid research. Nucleic acid dyes can bind to the bases in nucleic acid molecules, making the nucleic acids appear in a specific color or fluorescence. This change in color or fluorescence provides an intuitive means of observing the experimental results. By observing the stained nucleic acid samples, researchers can observe the morphology, size and direction of nucleic acids, and then understand the basic structural characteristics of nucleic acid molecules and the relative positions of different bases in the molecules. Nucleic acid dyes play an important role in the study of nucleic acid structure and function. By comparing the color or fluorescence changes of dyes under different conditions, researchers can infer the double helix structure of nucleic acid molecules, the base pairing rules, and the superhelical structure of DNA. In addition, dyes can also be used to analyze the degree of folding, sequence repetitiveness and other characteristics of nucleic acids, providing important information for a deeper understanding of the structure and function of nucleic acids.

[0003] Nucleic acid dyes also play a key role in nucleic acid detection technology. They can be used to detect, quantify and analyze nucleic acid samples, providing strong support for research in biology, medicine and other fields. With the deepening of research, the application of nucleic acid dyes in genetic testing, pathogen diagnosis and other aspects has gradually emerged, showing broad application prospects. In recent years, with the advancement of science and technology, nucleic acid dye technology has also been continuously innovating and developing. Some new nucleic acid dyes such as BIOTIUM nucleic acid dyes and SYBR series dyes have been introduced one after another. They have significant advantages in sensitivity, specificity, and ease of operation, bringing revolutionary changes to the field of nucleic acid detection. However, at the same time, controversies about the safety and stability of nucleic acid dyes have also continued to emerge. Therefore, how to ensure the safe use of dyes while ensuring the experimental effect has become a problem that needs to be solved urgently.

[0004] Licorice (Glycyrrhiza uralensis.Fisch.) is a commonly used Chinese medicine. Its roots and rhizomes are used as medicine. It has been accepted and used by people. It has detoxification, anti-inflammatory, antitussive, anti-tumor, anti-ulcer, antibacterial and other effects. Licorice has been recorded in Chinese herbal books of all dynasties. Licorice is not only a good medicine, but also known as the "king of all medicines". It has certain effects on the treatment of chronic bronchitis and asthmatic bronchitis. At the same time, glycyrrhizic acid is an organic compound. As a medicine, it has anti-inflammatory and anti-allergic effects; as a sweetener, it is widely used in various foods. Glycyrrhizic acid is a medicinal ingredient extracted from licorice. It has multiple effects and functions: Anti-inflammatory effect: Glycyrrhizic acid has certain anti-inflammatory effects and can relieve symptoms such as fever, cough, abdominal pain caused by inflammation. Liver protection effect: It has a certain therapeutic effect on liver inflammation caused by various reasons, can relieve liver inflammation, and promote the recovery of liver function. Regulating immunity: Glycyrrhizic acid can promote the regeneration and differentiation of immune cells, enhance the function of immune cells, and has a certain effect on regulating immunity, which can improve the body's resistance. In addition to the above effects, glycyrrhizic acid also has the effects of tonifying the spleen and replenishing qi. Glycyrrhizic acid mainly consists of two parts: glycyrrhetinic acid and glucuronic acid. Glycyrrhetinic acid has some unique physiological activities: Mineralocorticoid-like effect: Glycyrrhetinic acid can promote sodium and water retention, increase potassium excretion, and show a deoxycorticosterone-like effect. Glucocorticoid-like effect: The chemical structure of glycyrrhetinic acid is similar to that of corticosteroids, which can produce sodium retention and anti-diuretic and anti-inflammatory effects. Glucuronic acid is a naturally occurring compound with a wide range of application value: Anti-aging: Glucuronic acid has an antioxidant effect, which can neutralize free radicals and prevent them from damaging cells, thus having an anti-aging effect. Improve diabetes: Studies have shown that glucuronic acid can improve blood sugar control, reduce insulin resistance and postprandial blood sugar peaks, and reduce the risk of diabetic complications. Reduce the risk of heart disease: Glucuronic acid can reduce cholesterol levels, especially "bad" cholesterol, reduce inflammatory responses, and reduce the risk of heart disease. Beauty effect: Glucuronic acid can stimulate the production of collagen, make the skin more elastic, and reduce the production of dark spots and freckles. Glucuronic acid is a natural compound with many health and beauty benefits, and is widely used in the fields of medicine and beauty.

[0005] Carbon dots (CDs) are a class of zero-dimensional carbon nanomaterials with significant fluorescence properties, and their diameters are usually between 1-10 nm. They were first discovered in 2004 during the preparation of single-walled carbon nanotubes, and have gradually attracted attention in subsequent studies. As an emerging nanomaterial, CDs have shown great potential in scientific research and technological applications in recent years due to their unique physical and chemical properties, such as small size, large specific surface area, and high quantum efficiency. There are many different methods for the synthesis of CDs, mainly including "top-down method" and "bottom-up method". "Top-down method" includes arc discharge method, laser ablation method, electrochemical oxidation method, chemical exfoliation method, etc. These methods usually exfoliate nanocarbon dots from bulk carbon materials such as graphite and graphene. "Bottom-up method" uses carbon-containing small molecules such as glucose and citric acid to synthesize CDs through chemical reactions. In order to improve the quantum yield, photostability and adjust the emission wavelength of carbon dots, the synthesis process often includes passivation and doping steps. CDs have excellent fluorescence properties, their emission wavelength is adjustable, and their photostability is good. This makes CDs widely used in the fields of fluorescence display, bioimaging, optoelectronic devices, etc.; CDs have good biocompatibility and can be combined with biomolecules to form biosensors or used in drug delivery and other fields; CDs can maintain their structure and properties unchanged under various environmental conditions, which is crucial for their reliability in practical applications; CDs can be used in wastewater treatment, air pollution control and other fields to improve environmental quality by adsorbing harmful substances; CDs also show potential application value in quantum computing, optoelectronic devices and other fields. In recent years, with the in-depth understanding of the properties and application mechanisms of CDs, the research on CDs has made significant progress. In the future, with the continuous improvement of synthesis technology and the continuous development of new CDs, the application range of CDs will be broader. At the same time, in-depth research on the luminescence mechanism of CDs will also provide new ideas and methods for its application in the fields of photoelectric conversion, bioimaging and other fields. However, there has been no report on the application of CDs in the field of nucleic acid dyes. Summary of the invention

[0006] Nucleic acid dyes, as compounds that can bind to nucleic acids, have become a key tool for observing and analyzing nucleic acids in biological samples. They can help reveal the structure and function of nucleic acids and are an indispensable part of nucleic acid research. Although traditional nucleic acid dyes are widely used, they pose challenges to the environment and economy due to their high price, high toxicity, and long staining time. In order to overcome these problems, the present invention proposes a nucleic acid dye and its application in nucleic acid detection. The present invention uses licorice-derived carbon dots (CDs) as a new type of nucleic acid dye, which replaces traditional dyes for use in agarose gel electrophoresis to achieve efficient visual detection of DNA, RNA and plasmids.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] One of the technical solutions of the present invention:

[0009] A nucleic acid dye, which is licorice-derived carbon dots.

[0010] The present invention is the first to prepare carbon dots based on licorice root or licorice components as precursors, and successfully applies them to the field of nucleic acid detection. In the present invention, licorice root powder is first used as a biomass precursor in a system containing ethylenediamine to prepare a new type of CDs by a hydrothermal method. This CDs, as a nucleic acid dye, shows the potential for efficient visualization of DNA, RNA and plasmids in agarose gel electrophoresis. The experimental results show that the CDs are perfectly suitable for this field. Further, the present invention deeply studies glycyrrhizic acid, the main component of licorice, and successfully prepares new CDs using it as a biomass precursor in a system containing EDA. It is gratifying that this licorice-derived carbon dot-glycyrrhizic acid carbon dot can also be used in nucleic acid detection. Since glycyrrhizic acid is mainly composed of glycyrrhetinic acid and glucuronic acid, the present invention further uses these two components as precursors to prepare two new CDs. In a comparative experiment, it was found that glucuronic acid CDs showed better performance in nucleic acid detection. This discovery reveals that glucuronic acid in licorice root plays a key role in the preparation of CDs for nucleic acid dyes. The present invention not only provides a new biomass source for the preparation of CDs, but also provides a new efficient and visual method for nucleic acid detection, demonstrating the great potential and application value of licorice-derived carbon dots in the fields of nanomaterials and biology.

[0011] Preferably, the mass volume ratio of the licorice component to ethylenediamine is 1 g:300 μL.

[0012] Preferably, the method for preparing the licorice-derived carbon dots comprises the following steps: dissolving licorice root or licorice components and ethylenediamine in water, performing a hydrothermal reaction, and after the reaction is completed, cooling to room temperature, centrifuging, washing and drying the product to obtain the licorice-derived carbon dots.

[0013] Preferably, the licorice component includes glycyrrhizic acid, glycyrrhetinic acid or glucuronic acid.

[0014] Preferably, the temperature of the hydrothermal reaction is 180° C. and the time is 6 hours.

[0015] Preferably, when the raw materials of licorice-derived carbon dots are licorice root and ethylenediamine, the preparation method of licorice-derived carbon dots is: dissolve 1g of licorice root powder and 300μL of ethylenediamine in 10mL of deionized water, react at 180°C in a 50mL polytetrafluoroethylene-lined reactor for 6h, after the reaction is completed, cool to room temperature, centrifuge at 10000rpm for 15min, collect the supernatant, wash and filter through a microporous membrane (0.22μm), and vacuum freeze-dry the obtained suspension to obtain the licorice-derived carbon dots.

[0016] Preferably, when the licorice component is glycyrrhizic acid, the preparation method of licorice-derived carbon dots is as follows: 1 g of glycyrrhizic acid and 300 μL of ethylenediamine are dissolved in 10 mL of deionized water, reacted at 180° C. for 6 h in a 50 mL polytetrafluoroethylene-lined reactor, after the reaction is completed, cooled to room temperature, centrifuged at 10,000 rpm for 15 min, the supernatant is collected and washed and filtered through a microporous membrane (0.22 μm), and the obtained suspension is vacuum freeze-dried to obtain the licorice-derived carbon dots.

[0017] Preferably, when the licorice component is glycyrrhetinic acid, the preparation method of licorice-derived carbon dots is: dissolve 1g of glycyrrhetinic acid and 300μL of ethylenediamine in 10mL of deionized water, react at 180°C in a 50mL polytetrafluoroethylene-lined reactor for 6h. After the reaction is completed, cool to room temperature, centrifuge at 10000rpm for 15min, collect the supernatant, wash and filter through a microporous membrane (0.22μm), and vacuum freeze-dry the obtained suspension to obtain the licorice-derived carbon dots.

[0018] Preferably, when the licorice component is glucuronic acid, the preparation method of licorice-derived carbon dots is: dissolve 1g of glucuronic acid and 300μL of ethylenediamine in 10mL of deionized water, react at 180°C in a 50mL polytetrafluoroethylene-lined reactor for 6h, after the reaction is completed, cool to room temperature, centrifuge at 10000rpm for 15min, collect the supernatant, wash and filter through a microporous membrane (0.22μm), and vacuum freeze-dry the obtained suspension to obtain the licorice-derived carbon dots.

[0019] The second technical solution of the present invention:

[0020] The present invention also provides the use of the nucleic acid dye in nucleic acid detection. The licorice-derived carbon dots of the present invention can perform efficient visual detection of DNA, RNA and plasmids.

[0021] Preferably, the concentration of the licorice-derived carbon dots as nucleic acid dyes is 10-100 mg / mL.

[0022] More preferably, the concentration of the licorice-derived carbon dots as nucleic acid dyes is 100 mg / mL.

[0023] Compared with the difficulty in obtaining and safety of traditional nucleic acid dyes, the beneficial effects of the present invention are:

[0024] (1) The method of the present invention is simple to operate and has a fast synthesis speed. The entire preparation process of licorice-derived CDs does not involve complicated steps. In the system for synthesizing licorice-derived CDs, licorice roots or licorice components are reacted with ethylenediamine to obtain licorice-derived CDs.

[0025] (2) The binding mode between the nucleic acid dye prepared by the present invention and nucleic acid is different from the intercalation binding of traditional nucleic acid dyes. Traditional nucleic acid dyes are embedded in base molecules to cause mismatches, induce mutations, have potential carcinogenicity, cause harm to experimenters and pollute the environment. The binding mode between the licorice-derived CDs and nucleic acids of the present invention is groove binding. This binding mode will not cause problems such as base mismatches and is relatively safer than traditional nucleic acid dyes.

[0026] (3) The sensitivity of the nucleic acid dye prepared by the present invention is comparable to that of the traditional nucleic acid dye ethidium bromide, and can detect as little as 10 ng of the target.

[0027] (4) The novel nucleic acid dye prepared by the present invention has multiple advantages of low cost, easy synthesis and low toxicity, and has shown great potential in the field of nucleic acid detection, providing a research idea for promoting the further development and application of nanomaterials at the molecular level. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The drawings constituting a part of the present application are used to provide a further understanding of the present application. The illustrative embodiments and descriptions of the present application are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0029] Figure 1 A flow chart of the preparation and application of licorice-derived carbon dots CDs-4 provided in Example 4 of the present invention;

[0030] Figure 2 TEM and HRTEM images (inset) of the liquorice-derived carbon dots CDs-4 solution prepared in Example 4 of the present invention;

[0031] Figure 3 is a particle size distribution histogram of the liquorice-derived carbon dots CDs-4 solution prepared in Example 4 of the present invention;

[0032] Figure 4 The UV-visible spectrum, fluorescence spectrum and physical picture of the licorice-derived carbon dots CDs-4 prepared in Example 4 of the present invention, wherein "Day" represents daylight and "UV" represents ultraviolet light;

[0033] Figure 5This is a diagram showing the effects of different concentrations of licorice-derived carbon dots CDs-4 as nucleic acid dyes in Application Example 1;

[0034] Figure 6 This is a sensitivity effect diagram of DNA detection when the licorice-derived carbon dots CDs-4 in Application Example 2 are used as nucleic acid dyes;

[0035] Figure 7 This is a diagram showing the effect of licorice-derived carbon dots CDs-4 on the detection of DNA, RNA and plasmid.

[0036] Figure 8 The licorice-derived carbon dots CDs-1, CDs-2 and CDs-3 prepared in Examples 1-3 are used as nucleic acid dyes for the detection of DNA Marker III, wherein a is Example 1, b is Example 2, and c is Example 3. DETAILED DESCRIPTION

[0037] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0038] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0039] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0040] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0041] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0042] Unless otherwise specified, the room temperature in the present invention is 25±2°C.

[0043] All raw materials used in the examples of the present invention are commercially available.

[0044] The technical solution of the present invention is further illustrated by the following embodiments.

[0045] Example 1

[0046] 1g of licorice root powder and 300μL of ethylenediamine were dissolved in 10mL of deionized water and mixed thoroughly in a 50mL polytetrafluoroethylene-lined reactor. The reactor was reacted at 180°C for 6h. After the reaction was completed, it was cooled to room temperature and centrifuged at 10000rpm for 15min. The supernatant was collected and washed and filtered through a microporous membrane (0.22μm). The obtained suspension was vacuum freeze-dried to obtain licorice-derived carbon dots, which were recorded as CDs-1. It was in a powder state and was dispersed in ethanol for subsequent use.

[0047] Example 2

[0048] 1g glycyrrhizic acid and 300μL ethylenediamine were dissolved in 10mL deionized water and mixed thoroughly in a 50mL polytetrafluoroethylene-lined reactor. The reactor was reacted at 180°C for 6h. After the reaction was completed, it was cooled to room temperature and centrifuged at 10000rpm for 15min. The supernatant was collected and washed and filtered through a microporous membrane (0.22μm). The obtained suspension was vacuum freeze-dried to obtain licorice-derived carbon dots, which were recorded as CDs-2. It was in a powder state and was dispersed in ethanol for subsequent use.

[0049] Example 3

[0050] 1g of glycyrrhetinic acid and 300μL of ethylenediamine were dissolved in 10mL of deionized water and mixed thoroughly in a 50mL polytetrafluoroethylene-lined reactor. The reactor was reacted at 180°C for 6h. After the reaction was completed, it was cooled to room temperature and centrifuged at 10000rpm for 15min. The supernatant was collected and washed and filtered through a microporous membrane (0.22μm). The obtained suspension was vacuum freeze-dried to obtain licorice-derived carbon dots, which were recorded as CDs-3. It was in a powder state and was dispersed in ethanol for subsequent use.

[0051] Example 4

[0052] 1g of glucuronic acid and 300μL of ethylenediamine were dissolved in 10mL of deionized water and mixed thoroughly in a 50mL polytetrafluoroethylene-lined reactor. The reactor was reacted at 180°C for 6h. After the reaction was completed, it was cooled to room temperature and centrifuged at 10000rpm for 15min. The supernatant was collected and washed and filtered through a microporous membrane (0.22μm). The obtained suspension was vacuum freeze-dried to obtain licorice-derived carbon dots, which were recorded as CDs-4. It was in a powder state and was dispersed in ethanol for subsequent use.

[0053] The preparation and application flow chart of licorice-derived carbon dots CDs-4 provided in this example is shown in Figure 1 .

[0054] The transmission electron microscopy and high-resolution transmission electron microscopy images (inset) of the licorice-derived carbon dots CDs-4 provided in this example are shown in Figure 2 The particle size distribution histogram of the licorice-derived carbon dots CDs-4 provided in this embodiment is as follows: Figure 3 As shown, it can be seen that licorice-derived carbon dots were successfully prepared, and their particle size was 1.71±0.31nm.

[0055] The UV-visible spectrum, fluorescence spectrum and physical image of the licorice-derived carbon dots CDs-4 provided in this example are as follows: Figure 4 As shown, it can be seen that the excitation wavelength (Ex) and emission wavelength (Em) of CDs-4 prepared in Example 4 are 420nm and 496nm respectively; and the CDs-4 prepared in Example 4 exhibits strong absorption ability in the ultraviolet region of 250-300nm.

[0056] Application Example 1

[0057] The CDs-4 prepared in Example 4 was dispersed in ethanol at concentrations of 0.5 mg / mL, 1 mg / mL, 3 mg / mL, 5 mg / mL, 10 mg / mL, and 100 mg / mL, and used as a nucleic acid dye in agarose gel electrophoresis to detect nucleic acid products. The effect of CDs-4 prepared in Example 4 as a nucleic acid dye is shown in the figure Figure 5 As shown, it can be seen that the optimal concentration of CDs-4 prepared in Example 4 as a nucleic acid dye is 100 mg / mL.

[0058] Application Example 2

[0059] DNAMarkerⅢ (purchased from Shanghai Aladdin Reagent Company) was added to the sample wells at a sample volume of 1μL, 2μL, 3μL and 4μL at a voltage of 120V for 20min. After bromophenol blue was electrophoresed to 1-2cm of the gel, the staining effect of each band could be clearly observed in an automatic gel imaging analyzer (Bio-Rad Gel-Doc EZ USA). This was used to determine the minimum detection limit of CDs-4 prepared in Example 4 as a nucleic acid dye. The sensitivity effect diagram of CDs-4 prepared in Example 4 as a nucleic acid dye is shown in the figure below: Figure 6 As shown (Marker 3 is composed of a mixture of 7 double-stranded linear DNA fragments, the indicator band is 1200bp, and the rest are non-indicator bands, the target content is 50ng / 5μL, and the loading amount is controlled at 1μL, which can indicate that the amount of target contained in the remaining non-indicator bands is 10ng / μL), it can be seen that the CDs-4 prepared in Example 4 can detect 10ng / μL of DNA product.

[0060] Application Example 3

[0061] The CDs-4 prepared in Example 4 was used as a nucleic acid dye to detect DNA. Weigh 0.25g agarose, add 25mL 1XTAE (triacetate-acetic acid buffer), heat and wait for cooling to prepare 1w% agarose gel, add 100mg / mL CDs-4 solution, mix well, insert a comb and wait for solidification, then remove the comb and put it into the electrophoresis tank. The buffer solution must cover the gel. Add the obtained PCR product to the spotting well at a loading volume of 5μL and keep it at a voltage of 120V for 20min. After bromophenol blue is electrophoresed to 1-2cm of the gel, the corresponding band can be clearly seen on the gel imager. The effect diagram of DNA detection by CDs-4 prepared in Example 4 is shown in the figure. Figure 7 As shown, it can be seen that CDs-4 prepared in Example 4 can be used as a nucleic acid dye to detect the target DNA.

[0062] Application Example 4

[0063] The CDs-4 prepared in Example 4 was used as a nucleic acid dye to detect RNA. Weigh 0.25g agarose, add 25mL 1XTAE, heat and wait for cooling to prepare 1wt% agarose gel, add 100mg / mL CDs-4 solution, mix well, insert the comb and wait for solidification, then remove the comb and put it into the electrophoresis tank. The buffer solution must cover the gel. Add the obtained PCR product to the spotting well at a loading volume of 5μL and keep it at a voltage of 120V for 20min. After bromophenol blue is electrophoresed to 1-2cm of the gel, the corresponding band can be clearly seen in the gel imager. The effect diagram of CDs-4 prepared in Example 4 on RNA detection is shown in the figure. Figure 7 As shown, it can be seen that CDs-4 prepared in Example 4 can be used as a nucleic acid dye to detect the target RNA.

[0064] Application Example 5

[0065] The CDs-4 prepared in Example 4 was used as a nucleic acid dye to detect plasmids. Weigh 0.25g agarose, add 25mL 1XTAE, heat and wait for cooling to prepare 1wt% agarose gel, add 100mg / mL CDs-4 solution, mix well, insert a comb and wait for solidification, then remove the comb and put it into the electrophoresis tank. The buffer solution must cover the gel. Add the obtained PCR product to the spotting well at a loading volume of 5μL and keep it at a voltage of 120V for 20min. After bromophenol blue is electrophoresed to 1-2cm of the gel, the corresponding band can be clearly seen on the gel imager. The effect diagram of CDs-4 prepared in Example 4 on plasmid detection is shown in the figure below. Figure 7 As shown, it can be seen that CDs-4 prepared in Example 4 can be used as a nucleic acid dye to detect the target plasmid.

[0066] The detection effect of the licorice-derived carbon dots CDs-1, CDs-2 and CDs-3 prepared in Examples 1-3 as nucleic acid dyes on DNA Marker III is shown in FIG. Figure 8 , where a is Example 1, b is Example 2, and c is Example 3. It can be seen that licorice-derived carbon dots CDs-1, CDs-2, and CDs-3 can achieve efficient detection of DNA when used as nucleic acid dyes.

[0067] The above are only preferred specific implementations of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present application should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. A nucleic acid dye, characterized in that These are licorice-derived carbon dots.

2. The nucleic acid dye according to claim 1, characterized in that The preparation method of the licorice-derived carbon dots comprises the following steps: dissolving licorice roots or licorice components and ethylenediamine in water, carrying out a hydrothermal reaction, and after the reaction is completed, cooling to room temperature, centrifuging, washing and drying the product to obtain the licorice-derived carbon dots.

3. The nucleic acid dye according to claim 2, characterized in that The mass volume ratio of the liquorice root or liquorice component to ethylenediamine is 1 g: 300 μL.

4. The nucleic acid dye according to claim 3, characterized in that The licorice components include glycyrrhizic acid, glycyrrhetinic acid or glucuronic acid.

5. The nucleic acid dye according to claim 2, characterized in that The temperature of the hydrothermal reaction is 180° C. and the time is 6 hours.

6. Use of the nucleic acid dye according to any one of claims 1 to 5 in nucleic acid detection.

7. The use of the nucleic acid dye in nucleic acid detection according to claim 6, characterized in that: The concentration of the licorice-derived carbon dots as nucleic acid dyes is 10-100 mg / mL.

8. The use of the nucleic acid dye in nucleic acid detection according to claim 7, characterized in that: The concentration of the licorice-derived carbon dots as nucleic acid dyes is 100 mg / mL.

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