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A complete set of primers and methods for identifying the genetic relationship of Great Bustard

A kinship, great bustard technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as large damage, difficulty in collecting blood samples, and death

Active Publication Date: 2020-04-07
RES INST OF FORESTRY NEW TECH CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional blood collection can cause serious damage or even death, and it is difficult to collect blood samples for wild bustards
Non-damaged samples such as feces and / or feathers have been used for phylogenetic analysis in many endangered animals, but because the molecular markers used are species-specific, and the traditional PCR method is time-consuming and laborious, it is not as cost-effective as the multiplex PCR method. Therefore, There is an urgent need for a method for phylogenetic analysis and paternity identification of great bustards using non-invasive sampling

Method used

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  • A complete set of primers and methods for identifying the genetic relationship of Great Bustard
  • A complete set of primers and methods for identifying the genetic relationship of Great Bustard
  • A complete set of primers and methods for identifying the genetic relationship of Great Bustard

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1. Identifying the Genetic Relationship of the Great Bustard Based on Dual Molecular Markers of Mitochondrial DNA and Microsatellite DNA

[0077] Nine male great bustards and nine female great bustards were randomly selected from the Tumuji Nature Reserve in Inner Mongolia, and the naturally shed feathers or feces of each bustard were collected. Through the analysis of mitochondrial DNA and microsatellite dual molecular markers, the relationship between individuals can be judged. The 9 male bustards are numbered M1, M2, M3, M4, M5, M6, M7, M8, M9; the 9 female bustards are numbered F1, F2, F3, F4, F5, F6, F7, F8, F9.

[0078] 1. DNA extraction from feces or feathers

[0079] 1. The collected feces of the bustard were cryopreserved, and the genomic DNA of the bustard was extracted using a fecal DNA extraction kit (QIAamp DNAStool Mini Kit (ID: 51504) from QIAGEN); the isolated feathers of the collected bustard were stored in absolute ethanol , Genomic DNA of th...

Embodiment 2

[0114] Example 2. Verification of paternity identification of Great Bustard based on dual molecular markers of mitochondrial DNA and microsatellite DNA

[0115] Feces samples or feather samples of 2 adult male great bustards, 4 adult female great bustards and 2 chicks were collected from a free-range breeding farm (400m×300m) in Tumuji National Nature Reserve, Inner Mongolia, to identify the chicks. Biological father and biological mother. The two adult male bustards are numbered Male1 (abbreviated as Ma1, the same below) and Male2 (Ma2); the four adult female bustards are numbered Female1 (Fe1), Female2 (Fe2), Female3 (Fe3) and Female4 ( Fe4); the numbers of the two chicks are Nestling1 (Ne1) and Nestling2 (Ne2).

[0116] 1. DNA extraction from feces or feathers

[0117] 1. The collected feces of the bustard were cryopreserved, and the genomic DNA of the bustard was extracted using a fecal DNA extraction kit (QIAamp DNAStool Mini Kit (ID: 51504) from QIAGEN); the isolated f...

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Abstract

The invention discloses a primer set and method for identifying genetic relationship of great bustards. The primer set is composed of a primer set A and a primer pair capable of amplifying a great bustard mitochondrion DNA segment. The sequence of the great bustard mitochondrion DNA segment is disclosed as A1), A2) or A3): A1) nucleotide sequence disclosed as Sequence 23 in the sequence table; A2) A1)-derived DNA sequence with more than 90% of homogeneity to the DNA sequence defined in A1); or A3) A1)-derived DNA sequence hybridized with the DNA sequence defined in A1). The primer set A is composed of 20 single-strand DNAs disclosed as Sequence3-Sequence 20 in the sequence table. The experiment proves that the primer set and method can be used for identifying the genetic relationship among great bustards, and provide a technical support for provenance selection, pairing scheme establishment and genealogy construction in the great bustard artificial breeding and reintroduction process.

Description

technical field [0001] The invention relates to a set of primers and a method for identifying the genetic relationship of the Great Bustard in the field of biotechnology. Background technique [0002] The great bustard (Otis tarda), which belongs to the family Bustardidae of the order Hemiformes, is a bird of high international concern, and it is also a flagship species in the grasslands of China and an indicator species of grassland ecosystems. There are two geographical subspecies of Great Bustard, O.t.tarda and O.t.dybowskii (Palacín&Alonso, 2008). There are less than 1,000 species, more than half of which are fragmented in Northeast China and complete all stages of life history in China. Factors such as illegal hunting, habitat disturbance and overgrazing have led to the shrinkage of the distribution area and the sharp decline in the population of the oriental subspecies of the Chinese great bustard. Artificial breeding is one of the effective ways of ex situ conservat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143
Inventor 刘刚龚明昊崔丽娟胡德夫李林海韩莫日根孟德荣李惠鑫宁宇夏晓飞
Owner RES INST OF FORESTRY NEW TECH CHINESE ACAD OF FORESTRY