Detection method for identifying amycolatopsis
A technology of amycobacteria pseudomycoides and a detection method, which is applied in the field of primer pairs and detection for the detection of Amycolatopsis pseudomycoides, and can solve problems such as bacterial species identification that is unfavorable for a large sample size, lack of rapid detection methods, and long time consumption. , to achieve the effect of fast method, short time-consuming and good effect
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[0033] 1. Primer synthesis
[0034] Analyze the AMETH_3452 gene and its homologous sequences, and design PCR primers, see Table 1 for details. The primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. according to the sequence in the sequence listing.
[0035] Table 1
[0036]
[0037]
[0038] In the table, S=C or G; R=A or G; N=A, C, G or T; Y=C or T.
[0039] 2. Optimization of PCR reaction conditions
[0040] 2.1 The environmental wastewater DNA was used as a blank control, and the known DNA of A. methanolica 239 was used as a positive control. The bacterial strains were isolated and cultured, and the mycelium was directly scraped from the cultured petri dish, and the genomic DNA of the mycelium was extracted with a bacterial DNA extraction kit (Shanghai Jierui Bioengineering Co., Ltd.).
[0041] 2.2 Use the extracted DNA as a template, use the above primers, and use PCR to amplify. The annealing temperature of PCR was optimized by gradient PCR me...
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