Mercury ion detection method based on nucleic acid probe head-to-tail complementation strategy, and mercury ion detection kit based on nucleic acid probe head-to-tail complementary strategy

A detection kit and detection method technology, which can be used in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of low detection sensitivity, troublesome operation, increase experimental cost, etc., and achieve the effects of high sensitivity, simple operation and good specificity

Active Publication Date: 2015-10-14
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
View PDF2 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the potential disadvantage is that the detection sensitivity is low, and it is difficult to meet the needs of environmental sample detection, so it is necessary to improve the detection sensitivity through signal amplification
At present, tool enzymes (exonuclease, endonuclease, DNA polyme

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mercury ion detection method based on nucleic acid probe head-to-tail complementation strategy, and mercury ion detection kit based on nucleic acid probe head-to-tail complementary strategy
  • Mercury ion detection method based on nucleic acid probe head-to-tail complementation strategy, and mercury ion detection kit based on nucleic acid probe head-to-tail complementary strategy
  • Mercury ion detection method based on nucleic acid probe head-to-tail complementation strategy, and mercury ion detection kit based on nucleic acid probe head-to-tail complementary strategy

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0039] Example 1

[0040] A detection kit for mercury ions based on nucleic acid probe head-to-tail complementation strategy, including the following components:

[0041] (1) Streptavidin-modified magnetic beads;

[0042] (2) Biotin-modified DNA1, the sequence is as follows:

[0043] 5'-CAGTTTGTGTTTTCTCTTGC-Biotin-3' (SEQ ID NO. 1)

[0044] (3) DNA2, the sequence is as follows:

[0045] 5'-GCTTGAGATTTTCCATTCTGACTACTAGGGTCTGAGGG-3' (SEQ ID NO. 2)

[0046] (4) DNA3 and DNA4, the sequences are as follows:

[0047] DNA3: 5'-TACTCCCCCAGGTGCCCCTCAGACCCTAGTAGT-3' (SEQ ID NO. 3);

[0048] DNA4: 5'-GCACCTGGGGGAGTAACTACTAGGGTCTGAGGG-3' (SEQ ID NO. 4).

[0049] The 5' end of DNA3 is complementary to the 5' end of DNA4, the 3' end of DNA3 is complementary to the 3' end of DNA4, and the 3' end of DNA3 is complementary to the 3' end of DNA2;

[0050] (5) 20 mM Tris-acetate buffer (pH 7.5, containing 50 mM sodium acetate).

[0051] (6) Sybr Green I solution.

[0052] The working p...

Example Embodiment

[0057] Example 2

[0058] The detection method of mercury ions based on the head-to-tail complementation strategy of nucleic acid probes is carried out according to the following steps:

[0059](1) Add 1 mM biotin-modified DNA1 to the streptavidin-modified magnetic bead solution, mix well, react at room temperature for 30 minutes, separate the magnetic beads, and remove excess DNA1.

[0060] (2) The magnetic bead-DNA1 mixture was resuspended in 20 mM Tris-acetate buffer (pH 7.5, containing 50 mM sodium acetate), followed by the addition of 1 mM DNA2, and Hg 2+ , Mix well, react at room temperature for 30 minutes, separate magnetic beads, and remove excess DNA2.

[0061] (3) The above mixture was resuspended again with 20 mM Tris-acetate buffer (pH 7.5, containing 50 mM sodium acetate), then 2 mM DNA3 and 2 mM DNA4 were added, and the reaction was performed at room temperature for 60 minutes. Magnetic beads were separated to remove excess DNA3 and DNA4.

[0062] (4) The abo...

Example Embodiment

[0063] Example 3

[0064] for different concentrations of Hg 2+ detection of:

[0065] Formulated Hg 2+ Standard solutions at concentrations of 10 pM, 100 pM, 1 nM, 10 nM, 100 nM and 500 nM were stored at room temperature.

[0066] different concentrations of Hg 2+ The solution was respectively added to the reaction system described in Example 1, and the fluorescence intensity was detected after sufficient reaction, as shown in figure 2 shown, with Hg 2+ As the concentration increases, the corresponding fluorescence intensity also increases, when Hg 2+ When the concentration exceeds 100 nM, saturation is gradually reached. in Hg 2+ logarithm of concentration (lg 汞离子浓度 ) is the abscissa, and the fluorescence intensity is the ordinate. The standard curve is drawn. The two have a good linear relationship. The linear range is from 10pM to 100nM. The linear equation is: F = 191.8lg C + 18.4 (R=0.989) with a detection limit of 2 pM according to a 3x signal-to-noise ra...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention discloses a mercury ion detection method based on nucleic acid probe head-to-tail complementation strategy, and a mercury ion detection kit based on nucleic acid probe head-to-tail complementation strategy. According to the present invention, T base-rich nucleic acid is designed as a molecule recognition element, and is combined with a capture probe through T-Hg<2+>-T pairing so as to be immobilized on magnetic beads, two nucleic acid probes having head-to-tail complementation are added after the magnetic beads are separated, the long double-stranded DNA is formed through continuous hybridization complementation, a fluorescent intercalator Sybr Green I is added after the magnetic beads are separated, the fluorescent intercalator Sybr Green I has strong fluorescence property after being combined with the double-stranded DNA, and the fluorescence intensity and the mercury ion concentration have the good correlation so as to achieve the purpose of mercuric ion detection; the detection method and the detection kit have high sensitivity, the detection limit on Hg<2+> is 2 pM, the good specificity is provided, and other common interference ions do not affect the detection; and the signal amplification process is derived from the continuous head-to-tail complementation of the DNA probe without protease, the operation is simple, and the cost is low.

Description

technical field [0001] The invention belongs to the field of heavy metal ion detection, and relates to a mercury ion detection technology, in particular to a mercury ion detection method based on a nucleic acid probe head-to-tail complementary strategy and a detection kit. Background technique [0002] Mercury ions (Hg 2+ ) is a carcinogenic, teratogenic, and mutagenic environmental pollution source, which is a serious hazard to the ecological environment and human safety, and is an important indicator for environmental testing. Exposure to enrichment of mercury ions can lead to kidney failure, brain damage, nervous system and immune system damage, therefore, the effect on Hg 2+ Detection is important. The United States Environmental Protection Agency stipulates that the maximum allowable level of mercury ions in drinking water shall not exceed 10 nM. At present, the routine detection methods of mercury ions mainly include atomic absorption method, atomic fluorescence spe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/64
Inventor 陈俊华周顺桂
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap