Rice blast germ LAMP detection primer pairs and application
A technology for the detection of primers and rice blast fungus, applied in microorganisms, recombinant DNA technology, microorganism-based methods, etc., can solve problems such as late application, pesticide residue environment, and unsatisfactory disease prevention effect.
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[0020] 1. Design and screening of specific primers
[0021] (1) According to the gene sequence of Magnaporthe grisea Bt1 (accession number: XM_003718381), the online software PrimerExplorer was used to design the LAMP detection primer set, and 23 sets of primers were selected for specificity determination (Table 1).
[0022] Table 1 Part of the LAMP primer set designed according to the gene sequence of Magnaporthe grisea Bt1
[0023]
[0024] (2) 23 pairs of BtF3 / BtB3 primers were used to perform PCR amplification on the rice blast fungus genomic DNA. A total of 9 pairs of primers could amplify a single band from the rice blast fungus, and the 19th pair ( BtF3-19 (SEQ ID NO. 19) / BtB3-19 (SEQ ID NO. 24)) and the 22nd pair (BtF3-22 (SEQ ID NO. 25) / BtB3-22 (SEQ ID NO. 30)) primers ( figure 1 ).
[0025] 2. Specificity and sensitivity detection
[0026] (1) The results of qPCR showed that the primer pair BtF3-19 / BtB3-19 could detect the pathogen DNA after 24 cycles, while t...
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