Plateau zokor polymorphic microsatellite molecular markers

A molecular marker and microsatellite technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc.

Active Publication Date: 2021-12-03
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional microsatellite molecular marker development mainly adopts the magnetic bead enrichment method, which is time-consuming, laborious and expensive

Method used

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  • Plateau zokor polymorphic microsatellite molecular markers
  • Plateau zokor polymorphic microsatellite molecular markers
  • Plateau zokor polymorphic microsatellite molecular markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038]Example 1. Kit for analysis of genetic structure and genetic diversity of populations of plateau zokor and its close relatives

[0039] The components of the kit of the present invention include:

[0040] (1) PCR amplification reagent: comprising a pair of primers of SEQ NO:11-12, a pair of primers of SEQ NO:13-14, a pair of primers of SEQ NO:15-16, a pair of primers of SEQ NO:17-18, a pair of primers of SEQ NO:17-18, The primer pair of: 19~20, the primer pair of SEQ NO:21~22, the primer pair of SEQ NO:23~24, the primer pair of SEQ NO:25~26, the primer pair of SEQ NO:27~28, SEQNO: 29-30 primer pairs; (2) polymorphism detection reagent.

Embodiment 2

[0041] Example 2, Preparation of Polymorphic Microsatellite Molecular Markers in Plateau Zokor

[0042] (1) Extraction and detection of total genomic DNA

[0043] The total genomic DNA of the plateau zokor (coded as GY14) was extracted using Qiagen DNeasy Blood & Tissue Kit kit, and the extracted genomic total DNA was quantitatively and qualitatively detected by 1% agarose gel electrophoresis and Nanodrop 2000C spectrophotometer. The total DNA volume of the extracted genome was 50 μL. The electrophoresis results showed a bright main band with no obvious tailing. The spectrophotometer test results showed that the concentration was 67.3 ng / μL, and the OD260 / 280 was 2.12. The DNA quality met the requirements of simplified genome sequencing.

[0044] (2) Sequencing, clustering, assembly

[0045] The qualified plateau zokor genome total DNA samples were subjected to simplified genome sequencing using RAD-seq technology. Prepare a 300-700bp library and use the Illumina-HiSeqTM 250...

Embodiment 3

[0077] Embodiment 3, carries out plateau zokor genetic diversity analysis with described 10 pairs of primers

[0078] Genomic total DNA was extracted from the liver tissues of 29 plateau zokors from 7 different geographical populations and tested. The sample information is shown in Table 2. The 10 pairs of primers obtained by screening were synthesized with fluorescent primers, and the genomic DNA was qualified and diluted to 50 ng / μL for PCR amplification. The amplification system and reaction conditions were the same as those in Example 2. Fluorescent electrophoresis detection, and statistical data such as alleles according to the electrophoresis results.

[0079] The microsatellite locus polymorphism was evaluated on the electrophoresis results, and Popgene software was used to calculate the number of alleles (Na), effective number of alleles (Ne), observed heterozygosity (Ho) and expected heterozygosity (He), and PIC -CALC calculates the polymorphic information content (P...

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Abstract

The invention provides a group of plateau zokor polymorphic microsatellite molecular markers, and belongs to the technical field of DNA molecular markers. The microsatellite molecular marker(s) is / are a nucleotide sequence(s) as shown in any one or more of SEQ ID NO: 1 to SEQ ID NO: 10. Through high-throughput sequencing technology and by using a bioinformatics method, the invention screens microsatellite-containing DNA sequences, designs specific primers, and performs polymorphism detection on these microsatellite loci, and develop a group of (10) plateau zokor microsatellite molecular markers with abundant polymorphisms, and provides a set of plateau zokor microsatellite primers. The plateau zokor microsatellite primer pair provided by the invention can perform specific amplification on microsatellite loci, has high polymorphism, can be applied to the research fields of genetic diversity, population genetic structure, pedigree geography, evolution and genetic relationship and the like of the plateau zokor and related species thereof, and has excellent application prospects.

Description

technical field [0001] The invention belongs to the technical field of DNA molecular markers, in particular to a plateau zokor polymorphic microsatellite molecular marker. Background technique [0002] The plateau zokor (Eospalax baileyi) is a typical underground digging rodent, mainly distributed in the Qilian Mountains south of the Hexi Corridor in Gansu, the Gannan Plateau, the Qinghai Plateau, and the farmlands, hillsides, meadows, and plateaus of the northern and western highlands of Sichuan. Zokors have good adaptability to the natural conditions of the plateau and are endemic to the Qinghai-Tibet Plateau. Most of the behaviors of plateau zokor, such as foraging and reproduction, are carried out underground, often accompanied by high-energy-consuming excavation activities. Excessive excavation energy costs severely limit their long-distance migration activities, so the diffusion rate is significantly lower than that of above-ground animals. , it is of great significan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 蔡振媛张同作李文君宋鹏飞高红梅刘道鑫汪海静张婧捷江峰
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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