Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof

A biosensor, nucleic acid nanotechnology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of difficulty in practical application, difficulty in use and maintenance, expensive instruments, etc., to ensure detection sensitivity, rapid detection, The effect of easy preparation

Inactive Publication Date: 2012-02-15
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Mass spectrometry: Mass spectrometry is a common instrument used for detection. Although mass spectrometers have the characteristics of high sensitivity, less sample consumption, fast analysis speed, and multi-component analysis, the instruments are relatively expensive, and the use and maintenance are relatively expensive. difficulty
There is no such instrument in most areas, and it is difficult in practical application

Method used

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  • Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof
  • Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof
  • Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof

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Embodiment 1

[0030] Embodiment 1 Preparation of nucleic acid nano-gold biosensor of the present invention

[0031] 1. Design of six nucleic acid sequences

[0032] In existing Pb 2+ On the basis of enzymatically active DNAzyme, six sequences were specifically designed and synthesized as shown in SEQ ID NO: 1-6.

[0033] 2. Preparation of nano gold (colloidal gold):

[0034] Weigh 100g of 0.01% HAuCL4 solution into a 250ml round bottom flask, stir and heat until boiling; then quickly add 4ml of 1% trisodium citrate to the above solution, after the solution turns red, continue to boil for 10min, stop heating Continue stirring until cooling; the colloidal gold solution is stored at 4°C in the dark, and the nano-gold is identified by the maximum absorbance value at 520nm.

[0035] 3. Preparation of gold standard nucleic acid:

[0036] Dissolve the 1OD nucleic acid sequence in 100μl deionized water (take the third and fifth nucleic acid sequences, mix them in a molar ratio of 7:3, the con...

Embodiment 2

[0054] The nucleic acid nano-gold biosensor prepared in Example 1 was used to conduct the following experiments to verify its detection effect.

[0055] 1. Prepare lead ion standard solution gradients with concentrations of 100.0 μM, 50 μM, 10.0 μM, 1.0 μM, 500.0 nM, 100.0 nM, 0.0 nM, and store at room temperature.

[0056] 2. Prepare 10.0 μM Hg 2+ , Mn 2+ , Cd 2+ , Pb 2+ , Mg 2+ , Zn 2+ , Fe 2+ , Fe 3+ , Ba 2+ , Ca 2+ , Ni2+, Co 2+ solution.

[0057] 3. Take 1 tube of 1.0 OD nuclease and substrate with lead ion specificity, centrifuge at high speed for 10 seconds, slowly open the lid, add triple distilled deionized water, shake to dissolve, so that the final concentration is 100.0 μM ;

[0058] Mix the nuclease and substrate according to the ratio of 1:1.1, dilute with 4×SSC (pH8.0, replace NaCl with NaNO3), so that the final concentration of nuclease is 1μM, heat in a water bath at 70°C for 2 minutes, place in Cool slowly in warm water, this is the working fluid...

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Abstract

The invention discloses a nucleic acid nano-Au biosensor for detecting lead ions and a preparation method thereof. The nucleic acid nano-Au biosensor for detecting lead ions utilizes the principle that: when lead ions exist, the specific nuclease can cut the substrate which is complemented therewith, and utilizes the immune colloidal gold as an amplification color appearance system, and accordingly, the lead irons are quickly detected. The nucleic acid nano-Au biosensor for detecting lead ions has the advantages of high flexibility, low cost, no need of using any instrument and the like, not only the defect of requiring large-scale instruments in the former detection methods is solved, but also the detection flexibility can be ensured.

Description

technical field [0001] The invention relates to the field of metal ion detection, in particular to a nucleic acid nano-gold biosensor for detecting lead ions and a preparation method thereof. Background technique [0002] At present, the methods for detecting lead ions at home and abroad mainly include spectroscopic analysis, electrochemical analysis, mass spectrometry and so on. [0003] Spectral analysis methods include visible spectrophotometry, atomic absorption spectrometry, atomic fluorescence spectrometry, atomic emission spectrometry, and the like. Visible spectrophotometry has low sensitivity and low selectivity, and requires the use of highly toxic substances and volatile organic solvents, which have potential harm to experimenters and limit the application of this method. The experimental conditions of flame atomic absorption spectrometry and graphite furnace atomic absorption spectrometry are relatively complicated, and large-scale instruments are required. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/44
Inventor 曾令文方志远张文娟黄婧
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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