Pythium splendens braun fluorescence quantitative PCR detection reagent and detection kit and application
A technology that damps down bacteria and fluorescence quantification. It is used in fluorescence/phosphorescence, microbial determination/inspection, biochemical equipment and methods, etc. It can solve the problem that biological identification methods are labor-intensive and time-consuming, PCR sensitivity is not very high, and DNA cannot be detected. Samples and other problems, to achieve rapid and accurate qualitative and quantitative detection, reduce product contamination, and good repeatability
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[0026] Below in conjunction with embodiment the present invention is described in further detail.
[0027] 1. Design primers and probes
[0028] The present invention selects the conserved fragment in the ITS region of the oil palm damping-off pathogen as the target, which is reported in GenBank P . splendens The homology analysis and comparison of ITS sequences of similar species were carried out, and the conserved fragment (94bp) of the ITS region of oil palm damping-off was selected, and primers and probes were designed by using primer Express5.0 software. The synthesis of primers and probes adopts the chemical synthesis method of β-acetonitrile phosphorous acid amine, and the synthetic probe of OligoDNA is carried out by using an automatic DNA synthesizer, and the two ends of the synthesis are fluorescently labeled at the same time. The fluorescent reporter group labeled at the 5′ end of the probe is FAM, labeled at the 3' end with the non-fluorescent quencher TAMRA. A...
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