dsRNA and application thereof in pest control
A DNA molecule, insect technology, applied in the field of biological control, can solve problems such as hatching rate differences
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[0079] Embodiment 1, the synthesis of dsRNA and its application in the prevention and control of migratory locusts
[0080] 1. Synthesis of dsRNA
[0081] Using T7RiboMAX TM The Express RNAi System Kit synthesizes dsRNA. Specific steps are as follows:
[0082] 1) Preparation of recombinant plasmid
[0083] 1-1) Using migratory locust cDNA as a template, PCR amplification is performed with primers 1F / 1R and 2F / 2R to obtain a PCR product.
[0084] 1F: ACCTGTTGCTGCTTGATGTC;
[0085] 1R: GCCTTCTGTTTCTCGTCCTC.
[0086] 2F: AATGTCTGTCGTTCCTGCC;
[0087] 2R: TTCTCTCTCCTCCCTGCTGT.
[0088] The PCR reaction system is as follows (total volume 50 μL): cDNA template 1 μL, dNTP 4 μL, 10×Buffer 5 μL, pre-primer 1 μL, post-primer 1 μL, Taq enzyme 0.25 μL, ddH 2 O 38 μL.
[0089] The PCR reaction conditions are as follows: 95°C for 3min; 35 cycles of 95°C for 30s, 55°C for 30s, and 72°C for 1min and 30s; 72°C for 10min; and storage at 4°C.
[0090] 1-2) Perform electrophoresis of t...
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