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Microsatellite primer combination for identifying genetic relationship between eggs of Lophophorus lhuysii in same bird house and application

A green-tailed pheasant and microsatellite marker technology, applied in the field of molecular biology DNA molecular markers, can solve the problems of degradation, low DNA amount, etc., and achieve the effects of increasing accuracy, easy management, and good polymorphism

Active Publication Date: 2021-06-18
CHENGDU RES BASE OF GIANT PANDA BREEDING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA extracted from this non-destructive sampling sample has the problem of small amount of DNA and degradation, which also puts forward higher requirements for the screening of microsatellite markers

Method used

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  • Microsatellite primer combination for identifying genetic relationship between eggs of Lophophorus lhuysii in same bird house and application
  • Microsatellite primer combination for identifying genetic relationship between eggs of Lophophorus lhuysii in same bird house and application
  • Microsatellite primer combination for identifying genetic relationship between eggs of Lophophorus lhuysii in same bird house and application

Examples

Experimental program
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Effect test

Embodiment 1

[0040] In the present invention, microsatellite sites are developed in the whole genome sequence of the green-tailed iridescent pheasant, and primers for these sites are designed at the same time, and several nucleotide sequences containing microsatellite repeat sites are selected for species-specific screening. 200 pairs of species-specific primer sequences were synthesized as common primers, PCR was performed on 3 randomly selected DNA samples of the green-tailed iridescent pheasant, and electrophoresis was performed on agarose gel for primary screening of primers, and 93 microsatellite loci were obtained On this basis, 93 pairs of fluorescent primers were synthesized, PCR amplification and genotyping were carried out in multiple DNA samples, and the screening of microsatellite loci of the green-tailed rainbow pheasant was completed, and molecular markers with high polymorphism and good reproducibility were obtained 12 pairs. The specific content is as follows:

[0041] 1) ...

Embodiment 2

[0057] Using the QIAMP 69506 blood / tissue trace kit operation manual to extract 6 copies of DNA from eggshells (completed hatching or stillbirth) in 3 pens in 2018 in the captive population of the green-tailed rainbow pheasant in Fengtongzhai National Nature Reserve, and extracted Method is with embodiment 1. Twelve pairs of microsatellite primers were used (Table 1). The PCR reaction conditions were the same as in Example 1, and each reaction was repeated more than 3 times to ensure the consistency of the results.

[0058] Table 1 Primers

[0059]

[0060]

[0061] The above PCR products were genotyped by a first-generation gene sequencer, and the microsatellite typing data (amplified fragment length) are shown in Table 2.

[0062] Table 2 Microsatellite typing data (amplified fragment length bp)

[0063]

[0064] The genetic relationship of the above 6 samples was identified by software ML-Relate, and the analysis results are shown in Table 3. Among them, HS (Ha...

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Abstract

The invention discloses a microsatellite primer combination for identifying the genetic relationship between eggs of Lophophorus lhuysii in the same bird house and application. The primers comprise CM34, CM104, CM412, CM687, CM1423, CM1959, CM2014, CM2083, CM2269, CM2289, CM3131 and CM3590, and the nucleotide sequences of the primers are sequentially shown as SEQ ID NO.1-24. The primers can be stably amplified in the eggshell DNA of Lophophorus lhuysii, the microsatellite sequence of Lophophorus lhuysii with high polymorphism content and good repeatability is obtained through the 12 pairs of primers, the genotyping technology corresponding to the markers is established, and identification of three genetic relationship conditions of sib, half-sib or non-paternity between the eggs of Lophophorus lhuysii in the same bird house can be realized.

Description

technical field [0001] The invention belongs to the technical field of DNA molecular markers of molecular biology, and in particular relates to a combination of microsatellite primers and an application thereof for identification of phylogenetic relationship of eggs of a green-tailed rainbow pheasant. Background technique [0002] The green-tailed rainbow pheasant is a rare bird unique to my country. Like the giant panda, it was discovered and named in Baoxing County. The green-tailed rainbow pheasant is distributed in the inaccessible alpine areas of Sichuan, Yunnan, Qinghai, Gansu, and Tibet. It has extremely high ecological protection value and is also an umbrella species in the alpine meadow ecosystem. The number of green-tailed rainbow pheasant is rare in nature. In 2016, the expert team of IUCN estimated that the number of adults was between 2500 and 9999, and speculated that the wild population is still in the process of continuous decline. At present, the green-tail...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125C12Q2563/107Y02A50/30
Inventor 张亮侯蓉岳碧松沈富军吴里霞陈黎安俊辉张修月张文平寇洁王也李严刘佳文王涓刘红
Owner CHENGDU RES BASE OF GIANT PANDA BREEDING
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