Method for detecting uranium pollution by taking arabidopsis oligonucleotide fragment as probe
An oligonucleotide and Arabidopsis technology, which is applied in the field of molecular biology methods to detect environmental pollution, achieves the effects of specificity, specificity, accurate determination of pollution sources and whether the environment is polluted, and high sensitivity
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[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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