High-polymorphism single-copy microsatellite site combination and related primer combination for phoebe zhennan producing area tracing
A combination of primers and nanmu technology, applied in the biological field, can solve the problems of limited microsatellite sites, and achieve the effect of preventing false nanmu origin, wide application market, and good economic benefits
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Embodiment 1
[0060] Embodiment 1, the preparation of primer combination
[0061] This example provides a set of primer combinations that can be used for the evaluation of Phoebe genetic heterogeneity and origin traceability detection. The sequences of each primer are shown in Table 1. In Table 1, the one marked "F" is the forward primer, and the one marked "R" " is the reverse primer, and the 5' end of each forward primer is loaded with M13 sequence, that is, the 1-19th position of each forward primer. In the primer combination, each primer is packaged independently, and the molar numbers of each primer are equal.
[0062] Table 1. Primer sequences of 20 core microsatellite loci of Nanmu
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Embodiment 2
[0065] Embodiment 2, the detection of the origin of Phoebe
[0066] 1. Phoebe material:
[0067] A total of 124 Phoebe materials were selected from 4 populations in Guizhou, Chongqing, Hubei, and Sichuan, as shown in Table 2.
[0068] Table 2. Information on 124 Phoebe materials
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[0072] 2. Detection of the origin of Phoebe
[0073] Genomic DNA of 124 nanmu materials was extracted, and PCR amplification was performed using the primer combination in Example 1 using the genomic DNA of each nanmu material as a template.
[0074] A PCR amplification reaction system was prepared. For each system, a pair of primers, a genomic DNA, and a fluorescent dye were selected, and a fluorescent dye was selected for each pair of primers. Each genomic DNA was amplified separately using 20 primer pairs.
[0075] The PCR amplification reaction systems containing FAM, VIC, and NED are all as follows: 10×Buffer (Mg 2+ ) 1μL, 2mM dNTP 1μL, 5U / μL Taq DNA polyme...
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