Quarantine fruit fly multi-target rapid identification method
A multi-target, fruit fly technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effects of simple and easy steps, improved efficiency, and reduced quarantine costs
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Embodiment 1
[0050] Embodiment 1. The design of common primers for important economic fruit flies of Tephritidae
[0051] The mitochondrial genome sequences of all Diptera insect species were searched in the GenBank database, a total of 146 species.
[0052] The mitochondrial genome sequences of all 146 Diptera insect species obtained were compared with DNAMAN software.
[0053] The fruit flies of economically important fruit flies known from the mitochondrial genome sequences of 18 species (in the genera Anastrepha, Drosophila, Katrephus, Tephritus oligomanta, and Terephroides) The conserved loci that can specifically distinguish the important economic fruit flies from other closely related species such as Tephritidae and Diptera were screened.
[0054] For the conserved sites obtained by screening (such as B. dorsalis, the GenBank number of its mitochondrial genome sequence is DQ845759, and the positions of the conserved sites are 3142bp-3744bp), artificially design primers, use Oligo s...
Embodiment 2
[0057] Embodiment 2. Multi-target rapid identification method of important economic fruit flies
[0058] 1. DNA extraction:
[0059] Biological samples:
[0060] The 40 important economic fruit fly species in Table 1, including 15 fruit fly species with known mitochondrial genome sequences and 25 fruit fly species with unpublished mitochondrial genome sequences, are used to verify the generality of the specific primers of the present invention;
[0061] Procecidochares utilis, Drosophila melanogaster, and Adapsilia striatis in Table 1 are related species of Tephritidae and Diptera, and are used to verify the specificity of the specific primers of the present invention.
[0062] Use the "Blood / Cell / Tissue Genomic DNA Extraction Kit" to extract genomic DNA from a single head sample (both non-adult and adult worms are acceptable), and refer to the kit instructions for the specific operation process.
[0063] 2. Verification of primer generality and specificity:
[0064] Using ...
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