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Nucleic acid nano-gold biosensor used for detecting Hg2<+>

A biosensor and nucleic acid nanotechnology, applied in the field of biochemistry, can solve the problems of low detection sensitivity and high detection cost, and achieve low cost, high sensitivity, and easy preparation

Inactive Publication Date: 2010-10-06
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high specificity and can eliminate the interference of most ions in the actual sample system, but the detection sensitivity is not high, and DNA double labeling is required, and it also depends on fluorescence instruments, and the detection cost is still high

Method used

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  • Nucleic acid nano-gold biosensor used for detecting Hg2&lt;+&gt;
  • Nucleic acid nano-gold biosensor used for detecting Hg2&lt;+&gt;
  • Nucleic acid nano-gold biosensor used for detecting Hg2&lt;+&gt;

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1: Preparation of nucleic acid nano-gold biosensor of the present invention

[0023] 1. Design of three nucleic acid sequences

[0024] According to the technical principles of the present invention, the three nucleic acid sequences designed are:

[0025] Nucleic acid sequence 1: SEQ ID NO.1

[0026] 5’-thiol modification-ACACGCTAATCAAGCTTTAACTCATAGTTAGCGTGT-3’biotin modification

[0027] Nucleic acid sequence 2: SEQ ID NO.2

[0028] 5'-ACGCTTACTATGAGTTAAAGCTTG-3'

[0029] Nucleic acid sequence 3: SEQ ID NO.3

[0030] 5'-biotin modification-ACGCTAACTATGAGTTAAAGCTTGCTTAAG-3'

[0031] 2. Preparation of nano gold (colloidal gold):

[0032] 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...

Embodiment 2

[0044] Embodiment 2: Quality control and detection effect of the nucleic acid nano-gold biosensor of the present invention

[0045] The nucleic acid nano-gold biosensor prepared in Example 1 was used to conduct the following experiments to verify its detection effect.

[0046] 1. Preparation of Hg 2+ Standard solution gradient, concentrations are 1μM, 500nM, 100nM, 50nM, 20nM, 10nM.

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Abstract

The invention relates to a nucleic acid nano-gold biosensor used for detecting Hg2<+>, which comprises a detection test paper strip, a detection nucleic acid reagent and sampling buffer solution, wherein the detection test paper strip consists of a sample pad, a glass fiber, a nitrocellulose membrane and absorbent paper which are fixed on a rubber plate in turn from left to right; the glass fiber is coated with nano-gold labeled first nucleic acid; the nitrocellulose membrane is provided with a quality control line and a detection line; third nucleic acid is fixed on the quality control line, a 5' end of the sequence of the third nucleic acid is labeled with biotin which is fixed on the quality control line of the nitrocellulose membrane after reacting with streptavidin; the streptavidin is fixed on the detection line; the sequence of the third nucleic acid is complementary with the sequence of the first nucleic acid in specificity; and the detection nucleic acid reagent comprises second nucleic acid, and a sixth basic group starting from the 5' end has a mismatch of T-T basic groups with the sequence of the first nucleic acid. The nucleic acid nano-gold biosensor can quickly detect the Hg2<+>, needs no instrument, is convenient to use, and has high sensibility.

Description

technical field [0001] The invention belongs to the technical field of biochemistry and relates to a method for rapidly detecting Hg 2+ biosensors. Background technique [0002] At present, the main method for mercury detection is spectrometry, including atomic (absorption, emission, fluorescence, etc.) spectroscopic methods and spectrophotometry. In recent years, the combination of atomic spectroscopy and inductively coupled emission spectroscopy has improved the detection sensitivity and broadened the linear range of detection, while inductively coupled plasma mass spectrometry has made detection more sensitive and data analysis more convenient. However, such instruments are relatively expensive, which limits their widespread use in most laboratories. In addition to spectroscopy, chemical sensing methods represented by crown ether small molecules have also developed to a certain extent, but there are still some shortcomings such as low sensitivity and poor repeatability,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 曾令文黄婧顿博影方志远列浦昌萧卓
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI