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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

Pending Publication Date: 2021-10-08
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the microsatellite loci currently used for molecular markers of Phoebe are very limited. In order to effectively evaluate the genetic variation of Phoebe and accurately identify the source of species, a set of single-copy core microsatellite loci is urgently needed

Method used

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  • High-polymorphism single-copy microsatellite site combination and related primer combination for phoebe zhennan producing area tracing
  • High-polymorphism single-copy microsatellite site combination and related primer combination for phoebe zhennan producing area tracing
  • High-polymorphism single-copy microsatellite site combination and related primer combination for phoebe zhennan producing area tracing

Examples

Experimental program
Comparison scheme
Effect test

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

[0063]

[0064]

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

[0069]

[0070]

[0071]

[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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Abstract

The invention discloses a high-polymorphism single-copy microsatellite site combination and related primer combination for phoebe zhennan producing area tracing. The related primer combination for phoebe zhennan producing area tracing, disclosed by the invention, is composed of 40 single-stranded DNAs as shown in sequences 1-40 in a sequence table. By utilizing the primer combination provided by the invention, the genetic diversity of phoebe zhennan can be correctly evaluated, the origin of phoebe zhennan species can be quickly and accurately identified, and the primer combination is of great significance to protection of vulnerable species. Meanwhile, by utilizing the primer combination provided by the invention, the genetic composition of the phoebe zhennan can be conveniently determined, a phoebe zhennan fingerprint database can be conveniently established, the species source of the phoebe zhennan can be quickly and accurately identified, and a false phoebe zhennan producing area is prevented. The primer combination provided by the invention can be developed into a kit and made into a product, has a relatively wide application market, and good economic benefits are obtained.

Description

technical field [0001] The invention relates to the combination of high polymorphic single-copy microsatellite loci and related primer combinations in the field of biotechnology. Background technique [0002] Phoebe is a domestic second-class protected endangered species, endemic to my country, and a well-known traditional precious tree species with important social, cultural, and economic values. In history, Nan, Zhang, Catalpa, and Cui were collectively referred to as the four famous trees in the south of the Yangtze River, and Nanmu is at the top of it. "Emperor wood" was purchased and found in important buildings such as palace towers and temples in the Ming and Qing Dynasties, such as the Forbidden City and the Ming Tombs, where there are many Nanmu structures. There are many opinions on the name of "Phoebe": one kind of view thinks that Phoebe is the collective name of Phoebe species (China's main timber names, 2004; Li Donglin et al., 2004), and another kind of view ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 周世良张焱徐超
Owner INST OF BOTANY CHINESE ACAD OF SCI
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