Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for magnesium and sodium

A technology of amplification system and cleavage site, applied in the field of biosensors, can solve the problems of not being universally applicable, cumbersome and expensive sample processing, etc.

Inactive Publication Date: 2018-12-07
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Common detection methods for magnesium ions mainly include atomic spectrometry, including atomic absorption spectrometry, inductively coupled plasma emission spectrometry and fluorescence spectrometry, visible spectrophotometry, flow injection chemiluminescence and differential potential stripping, etc., but these The detection methods all require expensive instruments and professionally trained operators, and the sample pretreatment is also relatively cumbersome, which is not universally applicable

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  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for magnesium and sodium
  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for magnesium and sodium
  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for magnesium and sodium

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0152] Example 1 Magnesium and sodium cutting-type functional nucleic acid visual detection method for universal partition ultrafast amplification

[0153] 1. Experimental materials

[0154] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, sodium chloride, magnesium chloride, 2,2-azino-bis(3-ethane) Base-benzothiazole-6-sulfonic acid) diamine salt (ABTS), H 2 o 2 , 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), MES, LiCl, Tris, disodium edetate, tetramethylbenzidine, and urea were all purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.

[0155] The sequence design is as follows (SEQ ID NO:1-13):

[0156]

[0157] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;

[0158] The bold base sequence added to the end of the ribozyme substrate chain is to inc...

Embodiment 2

[0188] The specificity investigation of embodiment 2 detection method

[0189] According to the biosensor constructed in Example 1, 100nM Na + , 10μM Ag + , Mg 2+ , Zn 2+ 、Cd 2+ , Hg 2 + 、Cr 3+ Added to the system for detection, the results show that the established Na + The biosensor has good specificity ( Figure 8 ).

[0190] According to the biosensor constructed in Example 1, 50nM Mg 2+ , 5 μM Ag + 、Cu 2+ , Zn 2+ 、Cd 2+ , Hg 2+ 、Cr 3+ , Fe 2+ Added to the system for detection, the results show that the established Mg 2+ The biosensor has good specificity ( Figure 9 ).

[0191] According to the biosensor constructed in Example 1, 100nM Na + , 50nM Mg 2+ , 5 μM Ag + 、Cu 2+ , Zn 2 + 、Cd 2+ , Hg 2+ 、Cr 3+ , Fe 2+ Added to the system for detection, the results show that the established Mg 2+ The biosensor has good specificity ( Figure 10 ).

Embodiment 3

[0192] Embodiment 3 standard addition experiment

[0193] Get high-purity water and detect with the biosensor constructed in embodiment 1, Mg 2+ 、Na + No detection was carried out, and the standard addition experiment was carried out on it, and the results obtained by continuous measurement are shown in Table 1.

[0194] Table 1 The results of the standard recovery experiment

[0195]

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Abstract

The invention provides a universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for magnesium and sodium. According to the invention, primers, a template and probes are ingeniously designed, so ultrafast amplification of the template can be realized in the presence of magnesium and sodium ions, and amplification products are allowed to form G-quadruplexes in a proper environment; and color development is performed by utilizing the peroxidase-like activity of the G-quadruplexes. Thus, the problem of difficulty in realizing visual detection of traditional PCR products is overcome, and rapid and visual detection of magnesium and sodium ions is realized. Moreover, the method provided in the invention has the characteristics of high specificityand high sensitivity to magnesium and sodium ions, and is more objective and accurate in detection results.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a method for visually detecting magnesium- and sodium-cutting functional nucleic acids with universal partition ultrafast amplification. Background technique [0002] Sodium is a metal element, the chemical symbol is Na, the atomic number is 11, and the atomic weight is 22.9898. It belongs to Group IA and is a representative of alkali metal elements. It has a soft texture and can react with water to form sodium hydroxide and release hydrogen. Chemical properties Livelier. Sodium element is widely distributed in land and ocean in the form of salt, and sodium is also one of the important components in human muscle tissue and nerve tissue. The intake of sodium in the human body is mainly through intake of food salt. Without it, the diet will be tasteless and feel weak; but if the intake is too much for a long time, it will easily affect health and induce diseases, such as making...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851
CPCC12Q1/6851C12Q2531/113C12Q2525/301C12Q2525/186
Inventor 许文涛罗云波黄昆仑杜再慧田晶晶
Owner CHINA AGRI UNIV
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