Method for fast identifying frankliniella occidentalis
A Western flower thrips and a flower thrips technology are applied in the field of rapid identification of Western flower thrips to achieve the effects of good specificity and protection of the ecological environment
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Embodiment 1
[0020] ——Application of western flower thrips specific primers F.OL1 / F.OR for PCR identification
[0021] Total DNA was extracted from six species of thrips: Western flower thrips, flower thrips, yellow-breasted thrips, eight-section yellow thrips, tobacco thrips, and large thrips. / F.OR for conventional PCR amplification. The total volume of the PCR reaction is 50 μL, including: 10×PCR Buffer 5 μL, 25 mMol / L MgCl2 4 μL, 2.5 mMol / L dNTP 2 μL, Taq polymerase 0.3 μL (Takara, 5U / μL), 20 pmol F.OL1 / F.OR each 2 μL, template DNA 2 μL, and then add sterile water to stop the final volume of 50 μL. The reaction conditions are: 95°C for 5Min; 94°C for 50s, 53°C for 40s, 72°C for 50s, a total of 35 cycles; and finally 72°C for 6Min.
[0022] The PCR amplification products were detected by 1.5% agarose gel electrophoresis, and the detection results were as follows: figure 1 , after PCR amplification of the total DNA of Thrips occidentalis, the target band consistent with the expected f...
Embodiment 2
[0024] ——Application of western flower thrips specific primers F.OL2 / F.OR for PCR identification
[0025] Total DNA was extracted from six species of thrips: Western flower thrips, flower thrips, yellow-breasted thrips, eight-section yellow thrips, tobacco thrips, and large thrips. / F.OR for conventional PCR amplification. The total volume of the PCR reaction is 50 μL, including: 10×PCR Buffer 5 μL, 25 mMol / L MgCl2 4 μL, 2.5 mMol / L dNTP 2 μL, Taq polymerase 0.3 μL (Takara, 5U / μL), 20 pmol F.OL2 / F.OR each 2 μL, template DNA 2 μL, and then add sterile water to stop the final volume of 50 μL. The reaction conditions are: 95°C for 5Min; 94°C for 50s, 53°C for 40s, 72°C for 50s, a total of 35 cycles; and finally 72°C for 6Min.
[0026] The PCR amplification products were detected by 1.5% agarose gel electrophoresis, and the detection results were as follows: figure 1 , after PCR amplification of the total DNA of Thrips occidentalis, the target band consistent with the expected f...
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