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SSR fluorescence labeling primers for paternity test and population genetic analysis of Liangshantriton taliangensis

A paternity test and newt technology, applied in the field of genetics, can solve the problem of no microsatellite markers, etc., and achieve high observed heterozygosity, high data accuracy, and strong specificity

Active Publication Date: 2021-03-16
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there are no related reports on the application of microsatellite markers in paternity identification, population structure analysis and genetic management of Great Salamander

Method used

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  • SSR fluorescence labeling primers for paternity test and population genetic analysis of Liangshantriton taliangensis
  • SSR fluorescence labeling primers for paternity test and population genetic analysis of Liangshantriton taliangensis
  • SSR fluorescence labeling primers for paternity test and population genetic analysis of Liangshantriton taliangensis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Construction and acquisition of 10 pairs of primers

[0024] 1. Screening of SSR loci based on the simplified genome.

[0025]Take a salamander tail tissue sample, use the TIANGEN kit (TIANGEN Biotech Co., Ltd., Beijing), and follow the kit procedure for DNA extraction. 100 μL of DNA samples were obtained, and Illumina HiSeq2500 (PE250) sequencing technology was used for library construction. The specific steps of library construction (Novogene Biotechnology Co., Ltd.) include: (1) sample detection; (2) library construction and library inspection. Qualified samples were randomly interrupted with a Covaris ultrasonic breaker, and then repaired at the end and added with A Tailing, adding sequencing adapters, purification, PCR amplification and other steps to complete the entire library preparation. After the library construction is completed, use Qubit 2.0 for preliminary quantification, dilute the library, and then use Agilent 2100 to detect the insert fragm...

Embodiment 2

[0042] Example 2: Using primers to identify individuals of the Great Salamander

Embodiment 3

[0047] Example 3: Using primers for paternity identification of the Great Salamander

[0048] The 10 pairs of primers provided in Example 1 were used to carry out paternity identification on 30 progeny Great Salamanders (the female parents are all FGYHF34, and the candidate male parents include MGYHF31, MGYHF32 and MGYHF33). For the specific experimental process, refer to the PCR amplification and genotyping in Example 1. The typing results were read to obtain the locus matrix of 34 salamanders, and the probabilities of exclusion (PE) and cumulative PE (cumulative PE, CPE) of each locus were calculated using CERVUS v3.9 software. Use the Simulation of Parentage Analysis module for 10,000 simulated identifications, and the ParentageAnalysis module for formal parentage identification.

[0049] The relationship between the number of loci used and the probability of non-parental exclusion was sequentially tested according to the order of polymorphism (the combination of test prim...

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Abstract

The invention belongs to the field of genetics, and particularly relates to SSR fluorescence labeling primers for paternity test and population genetic analysis of Liangshantriton taliangensis. According to the specific technical scheme, a group of methods capable of performing individual identification, paternity test and population genetic structure analysis of the Liangshantriton taliangensis and 10 pairs of microsatellite primers are provided. The loci (primers) provided by the invention have the accuracy and polymorphism. When the technology provided by the invention is used for population selection research and population genetic analysis, scientific and feasible protection suggestions can be provided from the perspectives of propagation, genetic differentiation and the like, and a foundation is laid for future conservation work.

Description

technical field [0001] The invention belongs to the field of genetics, and in particular relates to SSR fluorescent-labeled primers for paternity identification and population genetic analysis of the great liang salamander. Background technique [0002] Liangshantriton taliangensis belongs to the family Salamandridae and belongs to the order Caudata. It is an endemic species in my country and is a second-class national key protected wild animal. It originally belonged to the genus Tylototriton, but Fei Liang (2012) divided it from the genus Tylototriton according to the morphological characteristics of its skull, and became the genus Liangshantriton independently. [0003] The great salamander has physiological and life history characteristics such as narrow distribution area, weak migration ability, and low reproductive potential, which make it very sensitive to environmental changes, and the population recovers slowly after decline. At present, due to ecological problems ...

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 谢锋束潇潇程明阳
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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