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Method for detecting lead by G-quadruplex fluorescence method formed by using unlabeled functional nucleic acid

A technology of functional nucleic acid and labeling function, which is applied in the direction of fluorescence/phosphorescence, biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of high sensitivity, high detection sensitivity and simple operation

Inactive Publication Date: 2013-09-18
SHANGHAI JIAO TONG UNIV
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  • Description
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Problems solved by technology

[0007] The present invention can overcome the defects in the prior art of needing to label functional nucleic acids and involving complex catalytic reactions, and has the characteristics of simple operation, high sensitivity, good selectivity, low cost and high efficiency

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  • Method for detecting lead by G-quadruplex fluorescence method formed by using unlabeled functional nucleic acid
  • Method for detecting lead by G-quadruplex fluorescence method formed by using unlabeled functional nucleic acid
  • Method for detecting lead by G-quadruplex fluorescence method formed by using unlabeled functional nucleic acid

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Embodiment 1

[0029] Such as figure 1 As shown, this embodiment includes the following steps:

[0030]1) Prepare the detection system formed by functional nucleic acid: In a 2mL graduated centrifuge tube, add 5 μL of the mother solution of functional nucleic acid (sequence: 5'-GGGTGGGTGGGTGGGT-3') with a concentration of 5 μM, add deionized water to 445 μL, and mix well Then place the centrifuge tube at 25°C for later use.

[0031] 2) Prepare a detection system with known lead concentration: get 16 centrifuge tubes containing the mixed solution of the detection system formed by functional nucleic acid prepared by step 1), add 50 μL lead standard solution of different concentrations respectively, so that the lead in the entire detection system Maintain the content at 0.5–600ppb, mix well, then place the centrifuge tube at 25°C and incubate for 10 minutes, add 5 μL of 400×DNA intercalator mother solution, incubate at 25°C in the dark for 10 minutes, and then reserve for the following determi...

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Abstract

The invention discloses a method for detecting lead by a G-quadruplex fluorescence method formed by using unlabeled functional nucleic acid, and belongs to the technical field of water quality testing. The method comprises the following steps that: a lead ion detection system is established through the functional nucleic acid and a DNA intercalating agent; if no lead ion exists in the detection system, the DN intercalating agent is intercalated into the functional nucleic acid, and fluorescence is released at 535 nm; after the lead ions are added into the detection system, the functional nucleic acid is promoted by the lead ions to form a G-quadruplex structure, and the DNA intercalating agent reacts with the G-quadruplex to suppress the release of the fluorescence at the 535 nm; on the base that a released fluorescence signal is in inverse ratio to the concentration of the lead ions, the lead ions can be quantitatively detected by determining the fluorescence signal. According to the method, the shortcomings a complex catalytic reaction is needed and the like because the functional nucleic acid is required to be labeled in the prior art are overcome; the method has the characteristics of simplicity in operation, high sensitivity, high selectivity, low cost and high efficiency.

Description

technical field [0001] The invention relates to a method in the technical field of water quality detection, in particular to a method for detecting lead by using a non-marked functional nucleic acid to form a G-quadruplex fluorescence method. Background technique [0002] Lead is a heavy metal element that seriously endangers human health. It is persistent, easy to migrate, and highly bioaccumulative. Lead entering the human body can damage the hematopoietic system, hinder the synthesis of hemoglobin, and cause anemia; damage the brain center and peripheral nervous system; affect the function of the digestive system, leading to anorexia; inhibit the synthesis and release of growth hormone, resulting in children's growth retardation; Absorption and metabolism of other metal elements; great damage to the kidneys, causing renal dysfunction; affecting the normal operation of the heart, causing myocardial damage, etc. For children, the academic community has confirmed that as lo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 周培詹深山吴远根罗艳芳詹学佳刘乐贺兰邢海波
Owner SHANGHAI JIAO TONG UNIV
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