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Application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg<2+> detection

A dual amplification and reaction technology, applied in the field of environmental pollutant detection, can solve problems such as increasing the complexity of probe design

Active Publication Date: 2015-12-09
SHANDONG UNIV
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  • Abstract
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Problems solved by technology

Although a two-step amplification is achieved, this process requires primers to generate a large number of DNA repeats and specific nicking enzyme recognition sites for nicking reactions, which increases the complexity of probe design

Method used

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  • Application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg&lt;2+&gt; detection
  • Application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg&lt;2+&gt; detection
  • Application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg&lt;2+&gt; detection

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Embodiment Construction

[0047] The present invention will be further described below in conjunction with embodiment.

[0048] Reagents and Instruments

[0049] The nucleic acid sequence was synthesized and purified by Sangon Bioengineering Co., Ltd. (Shanghai, China); the thermodynamic parameters and secondary structure of the nucleic acid sequence were simulated by the IDT online tool. Standard mercury solutions containing 2–5% nitric acid were purchased from Bailingwei Technology Co., Ltd. (Beijing, China). ExoIII was purchased from Thermo Fisher Scientific (Shanghai, China). Other drugs and reagents (all of analytical grade) were purchased from standard suppliers. All solutions were made of ultrapure water (>18.25MΩ·cm -1 ) preparation. Tap water was taken from the laboratory. Lake water was obtained from Baiyun Lake (Jinan, China). River water was taken from the Yellow River (Jinan, China). Sewage samples were taken from sewage treatment plants.

[0050] Hitachi F-7000 fluorescence spectr...

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Abstract

The invention discloses an application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg<2+> detection. When Hg<2+> exists, T-rich parts of H-DNA (deoxyribonucleic acid) and A-DNA form a T-Hg<2+>-T metal base pair, and H-DNA and A-DNA are hybridized to initiate a chain replacement reaction to form a Hg<2+>-mediated double-chain structure (referred as to Hg-Complex) containing an overhung protection part and a DNAzyme part. Under the action of Exo III, Hg<2+> and A-DNA are released and are hybridized with another H-DNA for cycle to obtain a large amount of DNAzyme sequences. The obtained DNAzyme sequences can cyclically cut molecular beacons to generate amplified signals. By means of effective combination of two cycles, Hg<2+> in an aqueous solution can be detected sensitively by means of this strategy, and high selectivity is realized. In the meantime, the method can be used for detecting Hg<2+> in sewage successfully and provides a potential tool for quantitative detection of Hg<2+> in an environment water sample.

Description

technical field [0001] The invention relates to the field of environmental pollutant detection, in particular to a dual amplification fluorescence reaction based on ExoIII (exonuclease III) assisted target cycle and DNAzyme (deoxyribozyme) cleavage cycle for detecting Hg 2+ , especially related to the detection of Hg in aqueous solution 2+ detection. Background technique [0002] Hg 2+ Mercury, the most common and stable form, is a serious environmental pollutant due to its highly toxic nature even at low concentrations. Because of its bioaccumulative properties, Hg 2+ ingestion is harmful to human health. When it exists in excess in the human body, it can damage the human nervous system, kidneys and other organs. Therefore, there is an urgent need to develop a reliable and sensitive method for Hg 2+ detection. Traditional detection methods including atomic absorption spectroscopy and inductively coupled plasma mass spectrometry have been developed for Hg 2+ detectio...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44
Inventor 姜玮王磊魏海平
Owner SHANDONG UNIV
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