Molecular marker method for cerambycidae larvae of different families
A molecular marker and pedigree technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of lack of larvae isolation and identification data, confusing identification, and undescribed larval morphology.
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[0026] 1. Test material:
[0027] The longicorn beetle specimens used were the specimens intercepted by the insect laboratories of Jiangsu Entry-Exit Inspection and Quarantine Bureau for many years. The sources and collection times of the five specimens are shown in Table 1. In addition, 28 beetle COI gene sequences registered in Genbank were selected for comparison, see Table 2.
[0028] Table 1 Name and source of experimental specimens
[0029]
[0030] Table 2 Genbank download sequence information
[0031]
[0032]
[0033] 2. Extraction of total genomic DNA
[0034] Using GenMag Animal Tissue / Cell Genomic DNA Extraction Kit: soak the worm body in alcohol, select muscle tissue of appropriate size, wash 2-3 times with sterile water, drain the water, grind the tissue, add appropriate amount of lysate Solution and proteinase K, 55 ℃ warm bath, so that the tissue is completely lysed. Then add magnetic beads and buffer to make the DNA adsorb to the magnetic beads, ...
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