Method for identifying molecule of athetis lepigone
A two-point identification technology of Spodoptera frugiperda and molecular identification, applied in the field of DNA sequence identification of insect species, can solve the problems of insufficient understanding of pests, mistaken for cutworms, and delays.
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Embodiment 1
[0028] 1. Collecting Insect Samples
[0029] In mid-to-early August, during the blooming period of the spodopteryx moth, adults were lured by lamps in 13 cities and counties in Hebei, Henan, Shandong, and Shanxi provinces. The typical characteristics of the adults were used to ensure that the sampled samples were spodoptera sp., and then Store in 95% ethanol at -20°C.
[0030] Table 1 The collection information of the population samples of Noctuid moth in the Second Committee
[0031] Table 1 Collecting data and individual number of Athelis lepigone
[0032]
[0033] 2. Extraction of Genomic DNA
[0034] Insect genomic DNA was extracted according to the Animal Tissue / Cell Genome Rapid Extraction Kit (SK8252) of Shanghai Sangon Bioengineering Co., Ltd., and stored at -20°C for later use.
[0035] 3. Amplification, cloning, sequencing and sequence analysis of COI gene
[0036] (1) Amplification of COI gene
[0037] Primers for PCR amplification of COI gene sequenc...
Embodiment 2
[0069] 1. Collecting Insect Samples
[0070] At the end of August, adults were lured by lamps in six cities and counties in Hebei Province. After being identified by experts according to the characteristics of the adults, they were placed in 95% ethanol and stored at -20°C.
[0071] Table 4 Sample collection information
[0072] Table 4 Collecting data of insects
[0073]
[0074] 2. Extraction of Genomic DNA
[0075] Insect genomic DNA was extracted according to the Animal Tissue / Cell Genome Rapid Extraction Kit (SK8252) of Shanghai Sangon Bioengineering Co., Ltd., and stored at -20°C for later use.
[0076] Amplification, cloning, sequencing and sequence analysis of COI gene
[0077] (1) Amplification of COI gene
[0078] Primers for PCR amplification of COI gene sequence: upstream primer / downstream primer 5-GGTCAACAAATCATAAAGATATTGG-3 / 5-TAAACTTGAGGGTGACCAAAAAATCA-3
[0079] 20 μL PCR reaction system: The total PCR amplification reaction system is 20 μL, i...
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