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Method for testing Pb2+ concentration by using DNA sensor based on G-quadruplex

A testing method and sensor technology, applied in the field of DNA sensors, can solve the problems of not avoiding DNA fluorescent labeling, complicated preparation process, unfavorable application, etc., and achieve the effect of meeting daily detection requirements, low cost, good sensitivity and selectivity

Inactive Publication Date: 2018-01-19
HENAN NORMAL UNIV
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Problems solved by technology

Based on this principle, he achieved the detection of Pb 2+ Although it has a good detection effect, the modification of DNA and the preparation process of GO are complicated and costly, which is also not conducive to practical application.
While Zhang et al. used Pb 2+ Pb 2+ For detection, although this method reached the detection limit of 0.4nM, it did not avoid the fluorescent labeling of DNA

Method used

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  • Method for testing Pb2+ concentration by using DNA sensor based on G-quadruplex
  • Method for testing Pb2+ concentration by using DNA sensor based on G-quadruplex
  • Method for testing Pb2+ concentration by using DNA sensor based on G-quadruplex

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Embodiment Construction

[0020] (1) Weigh NaCl, dissolve it with deionized water and dilute to 10mL, prepare to a concentration of 1M, store at 4°C, and dilute to the required concentration when used.

[0021] (2) Prepare a Tris-HCl buffer solution with pH = 6.0, then weigh 0.0169g of lead nitrate, dissolve it gradually with the prepared buffer solution and adjust the volume to 50mL, prepare it to a concentration of 1mM, and store it in the dark at 4°C , diluted to the desired concentration when used.

[0022] (3) Place Tris-HCl buffer (50mM, containing 50mM KCl, 10mM NaCl) containing G-quadruplex DNA (0.5μM) in a constant temperature water bath, keep it at different temperatures for 10 minutes, and then inject it In a 1cm quartz cuvette, the absorbance at 295nm was measured with a UV spectrophotometer.

[0023] (4) Put the mixed solution (containing 50mM Tris-HCl buffer solution pH6.0, 0.5μM DNA, different concentrations of KCl, 10mM NaCl, so that the total volume is 1mL) in a water bath at 95°C for...

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Abstract

The invention belongs to the technical field of a DNA sensor and specifically relates to a method for testing Pb2+ concentration by using the DNA sensor based on G-quadruplex. According to the invention, Pb2+ is detected only through a 'mixing-testing' step and the sensitivity and the selectivity are ultrahigh; compared with metallic ion auxiliary enzyme and RNA cracking deoxyribonuclease, the G-quadruplex DNA is more stable and no complex preparation process is required; compared with the scheme utilizing various shearing enzymes, the scheme is more economic and convenient; PPIX used as a fluorescent signal indicating unit is low-cost and has effective fluorescent characteristics; these mark-free characteristics can thoroughly reduce the test cost.

Description

technical field [0001] The invention belongs to the technical field of DNA sensors, in particular to a G-quadruplex-based DNA sensor for Pb 2+ Concentration test method. Background technique [0002] In aquatic ecosystems, lead ions are one of the most toxic environmental pollutants, which seriously threaten human life and health. Therefore, it is very important to develop sensors with ultra-high sensitivity and quantitative detection of lead ions. significance. However, traditional detection methods, such as atomic absorption spectrometry (AAS) or inductively coupled plasma mass spectrometry (ICP-MS), limit their use in on-site and real-time detection due to the expensive instrument itself and cumbersome pretreatment process. Through continuous efforts, researchers have made great progress in the detection method of lead ions. Some of these electrochemical detection methods have also been applied in practice, but their processing process is relatively complicated and req...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 王公轲邵超伟王双莉李丹闫长领陈得军
Owner HENAN NORMAL UNIV
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