Polyatomic element labeling probe and preparation method and application thereof
A technology for labeling probes and atomic elements, applied in biochemical equipment and methods, material inspection products, instruments, etc., can solve the problems of water solubility and stability attenuation, difficulty in controlling the polymerization degree of polymer chains, and difficulty in synthesizing element tags, etc. problem, to achieve the effect of simple purification process
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[0039]A 180bp long polyalkyne DNA backbone structure was synthesized by PCR, and the polyalkyne DNA was coupled to a macrocyclic metal ion complex containing azide group and Eu modification by click reaction to obtain polyatomic element-labeled probes.
[0040] Its synthesis process is as follows:
[0041] (1) Weigh the macrocyclic metal ion complex DOTA-N containing azide group and Eu modification 3 (Azido-mono-amide-DOTA, purchased from Macrocyclics (Dallas TX): B288) was dissolved in PBS buffer (pH=5.8), prepared into a 30mg / ml mother solution, and stored at 4°C;
[0042] (2) Take another 200 μL centrifuge tube, add DNA polymerase, DNA template, PCR buffer, A, G, C three conventional nucleotides and 5-ethynyl-dUTP, upstream and downstream primers (primer sequences are shown in Table 2 shown), magnesium sulfate, and add deionized water to make up to 50 μL. The DNA polymerase and the corresponding PCR buffer used are matching commercial polymerase and buffer (NewEngland Bio...
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