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Method for detecting uranium pollution by taking arabidopsis oligonucleotide fragment as probe

A technology of oligonucleotides and Arabidopsis thaliana, applied in the field of molecular biology detection of environmental pollution, to achieve high sensitivity, accurate determination of pollution sources and whether the environment is polluted, and specificity

Inactive Publication Date: 2011-09-28
HUNAN UNIV OF SCI & TECH
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to detect uranium pollution safely and accurately, and provide a method for detecting uranium pollution by using Arabidopsis thaliana-specific oligonucleotide fragments as a probe, which can keep away from the polluted environment Safe detection with high sensitivity and specificity

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  • Method for detecting uranium pollution by taking arabidopsis oligonucleotide fragment as probe
  • Method for detecting uranium pollution by taking arabidopsis oligonucleotide fragment as probe
  • Method for detecting uranium pollution by taking arabidopsis oligonucleotide fragment as probe

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Embodiment Construction

[0042] specific implementation

[0043] Production of Arabidopsis homologously up-regulated genes specifically expressed under uranium-induced induction of P. pinnata

[0044] 1. Collect the ghost needle grass in the uranium tailings area and the non-polluted control area, and store it in liquid nitrogen at -196°C for later use;

[0045] 2. Put 1 g of Devil's needles preserved in the refrigerator in a mortar and add liquid nitrogen to grind, and use the guanidine isothiocyanate method to extract total RNA (OD 260 / OD 280 equal to 1.8-2.0);

[0046] 3. Fluorescently label the sample RNA (Crystal Core cDNA Amplification Labeling Kit);

[0047] 3.1 Synthesis of 1-strand cDNA by reverse transcription

[0048] Start with Total RNA or mRNA, T7Oligo(dT)Primer containing T7 promoter sequence as primer, use CbcScript enzyme to synthesize 1-strand cDNA.

[0049] 3.2 Synthesis of 2-strand cDNA

[0050] Use RNase H to cut the RNA in the hybrid strand into short fragments, DNA Polyme...

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Abstract

The invention discloses a method for detecting Bidens maximowicziana Oett uranium pollution by taking an arabidopsis oligonucleotide fragment as a probe, comprising the steps of: hybridizing the total RNA (Ribose Nucleic Acid) extracted from the Bidens maximowicziana Oett in a uranium tailing area and a non-polluted contrast area with a 29KArabidopsis Genome Array chip to determine a specificallyexpressed up-regulated gene as the probe for detecting the uranium pollution; and hybridizing the total RNA extracted from the Bidens maximowicziana Oett in an environment to be detected and the non-polluted contrast area with the 29KArabidopsis Genome Array chip to detect whether a specifically expressed arabidopsis isogenous up-regulated gene exists or not, so as to know whether the Bidens maximowicziana Oett in the environment to be detected is polluted by the uranium or not. The detection method can perform safe detection far away from the polluted environment, solve the problem that the uranium pollution in the environment around the polluted area is difficult to detect correctly as well as correctly determine the pollution source and whether the environment is polluted or not, and has high sensitivity and undivided specificity of aiming at the uranium pollution only.

Description

technical field [0001] The invention relates to the technical field of detecting environmental pollution by molecular biology methods, in particular to a method for detecting uranium pollution of P. pinnata by using Arabidopsis thaliana specific oligonucleotide fragments as probes. Background technique [0002] The traditional uranium pollution and the detection of radioactive substances in the environment are usually detected by radioactive detectors, and the probes of α, β, χ, γ multifunctional ray detectors are used to detect the radiation dose. The disadvantages of this detection method are: (1) Although it can detect a certain dose of radioactive sources, it is difficult to distinguish what kind of radioactive substances; (2) its sensitivity has certain limitations; (3) the detection must be close to the polluted environment, which poses a threat to the health of the inspectors . At present, the method of detecting environmental uranium pollution by molecular biology m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 肖璐高利臣冯涛
Owner HUNAN UNIV OF SCI & TECH
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