Matched reagent for identifying aedes-aegypti drug resistance and application thereof
A technique for Aedes mosquitoes and reagents, which is applied to a complete set of reagents for identifying drug-resistant traits of Aedes aegypti and its application field
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Embodiment 1
[0064] Embodiment 1, the preparation of the complete set of reagents of identification Aedes aegypti drug-resistant character
[0065] A complete set of reagents for identifying drug-resistant traits of Aedes aegypti, consisting of primer sets named A and B respectively;
[0066] A is composed of CD1, CD2 and CD3, B is composed of CD1, CD2 and CD4, CD1 is the single-stranded DNA shown in SEQ ID No.1, CD2 is the single-stranded DNA shown in SEQ ID No.2, and CD3 is SEQ ID The single-stranded DNA shown in No.3, CD4 is the single-stranded DNA shown in SEQ ID No.4.
[0067] CD1 and CD2 can amplify the 587bp DNA fragment shown in sequence 5 from the wild-type Aedes aegypti genome or the Aedes genome containing the V1016G mutation, and CD2 and CD3 can be amplified from the wild-type Aedes aegypti genome The 344bp DNA fragment shown in sequence 6, CD2 and CD4 can amplify the 344bp DNA fragment shown in sequence 7 from the Aedes aegypti genome containing the V1016G mutation.
Embodiment 2
[0068] Embodiment 2, Aedes aegypti drug resistance detection
[0069] Genomic DNA of 100 Aedes aegypti mosquitoes numbered AA001-AA100 captured in the field were extracted respectively.
[0070] The genomic DNA of each Aedes aegypti was subjected to PCR amplification according to the following method (amplification system is shown in Table 1):
[0071] (1) Take reagent T (Treasure Bioengineering Co., Ltd., product number DR011, the components of reagent T are shown in Table 2) and add them to 2 PCR tubes, labeled tube A and tube B, 12.5 μL in each tube.
[0072] (2) Add 1.1 μL of reagent P1 to tube A, and add 1.1 μL of reagent P2 to tube B. Reagent P1 is the solution that CD1, CD2 and CD3 of embodiment 1 and water form, and the molar ratio of CD1, CD2 and CD3 is 3:4:4 in the reagent P1; Reagent P1 is the CD1 of embodiment 1, CD2 and CD4 and water The solution composed, the molar ratio of CD1, CD2 and CD4 in reagent P1 is 3:4:4.
[0073] (3) Add 9.9 μL deionized water to tub...
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