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Visual detection method of magnesium and zinc cutting type functional nucleic acids for universal partition and ultra-fast amplification

A technology of amplification system and cleavage site, applied in the field of biosensors, which can solve the problems of complex pretreatment, long detection period, high price, etc.

Active Publication Date: 2018-11-20
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional magnesium and zinc ion detection methods can generally be divided into cold atomic absorption spectrometry, graphite carbon atomic absorption spectrometry, and flame atomic absorption spectrometry, etc., but there are common problems such as complex pre-treatment, large-scale instrument operation, long detection cycle, and high price. Characteristics

Method used

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  • Visual detection method of magnesium and zinc cutting type functional nucleic acids for universal partition and ultra-fast amplification
  • Visual detection method of magnesium and zinc cutting type functional nucleic acids for universal partition and ultra-fast amplification
  • Visual detection method of magnesium and zinc cutting type functional nucleic acids for universal partition and ultra-fast amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1 Magnesium and zinc-cut functional nucleic acid visual detection method for ultrafast amplification of universal partition

[0144] 1. Experimental materials

[0145] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, magnesium chloride, zinc chloride, 2,2-azino-bis(3-ethane) Base-benzothiazole-6-sulfonic acid) diamine salt (ABTS), H 2 o 2 , 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), sodium hydroxide, and disodium hydrogen phosphate were all purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.

[0146] The sequence design is as follows (SEQ ID NO:1-11):

[0147]

[0148] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;

[0149] The bold base sequence added to the end of the ribozyme substrate chain is to increase the Tm value of bind...

Embodiment 2

[0178] Embodiment 2 Specificity investigation of detection method

[0179] According to the biosensor constructed in Example 1, 1 μM Mg 2+ , 500nM Zn 2+ , 10μM Ag + 、Cu 2+ 、Cd 2 + , Hg 2+ 、Cr 3+ , Fe 2+ , Ca 2+ , added to the system for detection, the results show that the established Mg 2+ , Zn 2+ The biosensor has good specificity ( Figure 8 ).

Embodiment 3

[0180] Embodiment 3 Standard addition experiment

[0181] Get high-purity water and detect with the biosensor constructed in embodiment 1, Mg 2+ , Zn 2+ If there is no detection, the standard addition experiment is carried out on it, and the results obtained by continuous measurement are shown in Table 1 and Table 2.

[0182] Table 1 The results of the standard recovery experiment

[0183]

[0184] Table 2 The results of the standard recovery experiment

[0185]

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Abstract

The invention provides a visual detection method of magnesium and zinc cutting type functional nucleic acids for universal partition and ultra-fast amplification. The visual detection method providedby the invention can perform ultra-fast amplification on templates the presence of magnesium and zinc ions by ingeniously designing primers, the templates and probes, enables amplification products toform a G-quadruplex in a suitable environment, further uses peroxidase-like activity of the G-quadruplex for color development, solves a problem that traditional PCR products are hard to be detectedvisually, and realizes fast and visual detection of magnesium and zinc ions. Moreover, the visual detection method provided by the invention has the characteristics of high specificity and high sensitivity to magnesium and zinc ions, and more objective and accurate 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 zinc-cutting functional nucleic acids with universal partition ultrafast amplification. Background technique [0002] Magnesium, the chemical symbol is Mg, the atomic number is 12, and the atomic weight is 24. It belongs to Group IIA. It is a silver-white metal with metallic luster and is widely distributed in nature. The Chinese Nutrition Society recommends that adult men need about 350 mg of magnesium per day, adult women need about 300 mg, pregnant women and breastfeeding women need about 450 mg, children aged 2-3 need 150 mg, and children aged 3-6 need 200 mg. The tolerable upper intake level (UL) was set at 700mg / d. Magnesium is the main cation in human cells. It is concentrated in mitochondria, second only to potassium and phosphorus, and ranks third in extracellular fluid after sodium and calcium. It is an essential substan...

Claims

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Application Information

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IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2521/337C12Q2525/301C12Q2565/607
Inventor 许文涛罗云波黄昆仑杜再慧田晶晶
Owner CHINA AGRI UNIV
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