Identification method and special primer for trogoderma glabrum and trogoderma variabile
The technology of the black beetle and the black beetle, which is applied in the biological field, can solve the problems of no rapid molecular biology detection method, time-consuming and high cost, shorten the time required for the detection, improve the detection accuracy, and achieve credible results. high degree of effect
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Embodiment 1
[0042] Embodiment 1, the design of primer
[0043] According to the sequence (gene sequence number: FJ58973) of the existing Trogoderma zebra, design primers:
[0044] Tgla-F: 5-CAACATTTATTTTTGATTT-3 (SEQ ID NO: 1)
[0045] Tgla-R: 5-TCCAATGCACTAATCTGCCA-3 (SEQ ID NO: 2)
Embodiment 2
[0046] Embodiment 2, identification of Dermoderma melanogaster and Dermoderma versicolor
[0047] 1. Extraction of DNA from dermatomid samples
[0048] The two dermatomid beetles numbered 1 and 2 were respectively provided by Huangpu Entry-Exit Inspection and Quarantine Bureau (collected in Huangpu, Guangzhou) and Shandong Agricultural University (collected in Tai’an, Shandong). Tiangen Tissue Genomic DNA Extraction Kit (TIANamp Genomic DNA Kit ), follow the instructions to extract the total DNA of the sample to be tested. Specifically: Grind live worms in 200 μl of GA buffer, mix well, add 20 μl of proteinase K, and place at 37° C. for 4 hours. Then add 200 μl of GB buffer, mix well, and place at 70° C. for 10 minutes. Add 200ul absolute ethanol and mix well for 15s. Transfer the obtained solution into the CB3 adsorption column, centrifuge at 12,000rpm for 30s, pour off the waste liquid, and put the adsorption column back into the collection tube. Add 0.5ml of GD buffer s...
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