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Mongolian wild donkey microsatellite molecular marker combination as well as primer and application of Mongolian wild donkey microsatellite molecular marker combination

A technology of molecular markers and microsatellites, applied in biochemical equipment and methods, measurement/testing of microorganisms, recombinant DNA technology, etc., can solve the problems of few molecular genetics studies, threat to the survival of Mongolian wild donkeys, etc., and achieve repeatability Good results

Pending Publication Date: 2022-08-02
XINJIANG INST OF ECOLOGY & GEOGRAPHY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to agricultural land use, overgrazing and economic development, the habitat of the Mongolian wild ass is gradually fragmented and marginalized, resulting in the survival of the Mongolian wild ass still being threatened and endangered
At present, the domestic research on the Mongolian wild ass mainly focuses on the ecological population structure, behavior mechanism and habitat, and there are very few related studies on its molecular genetics

Method used

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  • Mongolian wild donkey microsatellite molecular marker combination as well as primer and application of Mongolian wild donkey microsatellite molecular marker combination
  • Mongolian wild donkey microsatellite molecular marker combination as well as primer and application of Mongolian wild donkey microsatellite molecular marker combination
  • Mongolian wild donkey microsatellite molecular marker combination as well as primer and application of Mongolian wild donkey microsatellite molecular marker combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of Mongolian wild donkey polymorphic microsatellite molecular markers

[0050] 1. Extraction of blood genomic DNA

[0051] Using (Qiagen) DNeasy Blood & Tissue Kit (50) kit, the blood genomic total DNA of Mongolian wild donkey was extracted. The total genomic DNA extracted was quantitatively and qualitatively detected by agarose gel electrophoresis and ultraviolet spectrophotometer. Qualified Mongolian wild donkey blood genomic DNA was sequenced by Illumina PE150. After sequencing alignment, screening filtering, quality assessment and assembly, an effective data volume (Clean Base) of 71571.96Mb was obtained, the Effective Rate (Effective Rate) was 99.63%, the base error rate was 0.03%, the Q20 was 97.19%, and the Q30 was 92.38%, and the GC content was 42.81%. It indicates that the data volume of the sample is sufficient, the sequencing quality is high (Q20≥95%, Q30≥90%), the GC distribution is normal, and the library construction and sequencing...

Embodiment 2

[0065] Example 2 Polymorphism identification and population genetic diversity analysis of Mongolian wild donkey microsatellite molecular markers

[0066] 1. Fluorescent primer synthesis was performed on 8 pairs of primers corresponding to the 8 microsatellite sites screened in Example 1, that is, fluorescent labeling (FAM-blue) was performed on the 5' end of the single-sided primer; constructed in Example 1 The fecal DNA of 80 parts of Mongolian wild ass in the Mongolian wild ass DNA template library was a template, and PCR amplification was carried out, and the amplification system and reaction conditions were the same as in Example 1, and the amplified product was in an ABI 3730XL sequencer (Shanghai Jierui Bioengineering). Co., Ltd.) performed capillary fluorescence electrophoresis detection, and counted data such as alleles according to the electrophoresis results.

[0067] The collected data were separated by fluorescence color using GeneMarker v2.2.0 (SoftGenetics LLC., ...

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Abstract

The invention discloses a Mongolian wild donkey microsatellite molecular marker combination as well as a primer and application thereof, and relates to the technical field of DNA molecular markers. The Mongolian donkey microsatellite molecular marker combination comprises a nucleotide sequence Eqr3400, a nucleotide sequence Eqr5948, a nucleotide sequence Eqr732, a nucleotide sequence Eqr166, a nucleotide sequence Eqr1096, a nucleotide sequence Eqr1226, a nucleotide sequence Eqr652 and a nucleotide sequence Eqr821, wherein the nucleotide sequences of the nucleotide sequences are shown as SEQ ID NO. 17-24. An amplification primer combination of the molecular marker combination comprises nucleotide sequences as shown in SEQ ID NO. 1-16. The invention develops a group of Mongolian wild donkey microsatellite molecular markers with abundant polymorphism, and provides a group of Mongolian wild donkey microsatellite primers. The Mongolian wild donkey microsatellite primer pair provided by the invention can be used for carrying out specific amplification on microsatellite sites and has high polymorphism.

Description

technical field [0001] The invention relates to the technical field of DNA molecular markers, in particular to a Mongolian wild donkey microsatellite molecular marker combination, primers and applications thereof. Background technique [0002] Microsatellite: Microsatellite (simple sequence repeat, SSR) refers to a DNA simple tandem repeat sequence, which consists of a core sequence and flanking sequences on both sides. The core sequence is 1-6 nucleotide repeat units. The difference in the number of repeats and repeat units between different alleles makes it polymorphic in length. Both sides of the core sequence are composed of conserved flanking sequences, which are common among species. Higher homology, so it can be used for primer design. Compared with other molecular markers, microsatellites have the advantages of large number, good stability, rich polymorphism information, co-dominance and easy detection. At the same time, microsatellite analysis based on PCR technol...

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 汪沐阳丁佳佳梅宇徐文轩杨维康师玮
Owner XINJIANG INST OF ECOLOGY & GEOGRAPHY CHINESE ACAD OF SCI