LAMP detection primer for detecting C. gloeosporioides in olive tissue
A technology for detecting anthracnose bacteria and primers, which is applied in the field of plant pathology, can solve problems such as inability to truly and accurately classify pathogenic bacteria, and achieve the effects of guaranteed reliability of results, strong practicability and good practicability.
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[0024] 1 Extraction of mycelium genomic DNA:
[0025] Genomic DNA was extracted according to the instructions of the high-efficiency plant genomic DNA extraction kit. DNA purity was checked using a spectrophotometer.
[0026] 2 Design of LAMP-specific primers
[0027] The translation elongation factor alpha 1 gene (TEF1) gene, which is highly conserved in gene and expression regulation in different species, was selected as the target gene. Find and download the TEF1 gene sequence of G. anthracnose and its related species from the NCBI website, use BioEdit software for sequence comparison, analyze and find the specific conserved sites of G. anthracis, and then use PrimerExplorer V5 (http: / / primerexplorer.jp / lampv5e / index.html) online software to design primers (Table 1).
[0028] Table 1 LAMP primers and sequences
[0029]
[0030] 3 Establishment of LAMP amplification detection system
[0031] Using the DNA extracted in (1) above as a template, use the outer primer F3...
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