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Hyla tsinlingensis microsatellite sites, primers for microsatellite sites and application of microsatellite sites and primers

A technology of microsatellite sites and hyla, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as blanks

Inactive Publication Date: 2015-02-04
ANHUI UNIVERSITY
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However, research in this area is still blank

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  • Hyla tsinlingensis microsatellite sites, primers for microsatellite sites and application of microsatellite sites and primers
  • Hyla tsinlingensis microsatellite sites, primers for microsatellite sites and application of microsatellite sites and primers
  • Hyla tsinlingensis microsatellite sites, primers for microsatellite sites and application of microsatellite sites and primers

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Embodiment Construction

[0008] The present invention will be described in detail below in conjunction with specific embodiments.

[0009] 1. DNA extraction

[0010] Genomic DNA of Hyla Qinling was extracted using muscle or blood tissue. Genomic DNA was extracted using SDS / proteinase K cleavage and phenol-chloroform extraction (Sambrook et al., 1999).

[0011] 2. Construction of microsatellite enrichment library of Hyla Qinling

[0012] Genomic DNA was extracted, and AFLP rapid isolation method (Fast Isolation by AFLP of Sequences Containing repeats, FIASCO) was used to construct a microsatellite enrichment library. The specific process was as follows: Take about 250ng of genomic DNA, digest it with MseI, and at the same time combine it with the AFLP receptor of MseI Connector (5'-TAC TCA GGA CTC AT-3' / 5'-GAC GAT GAG TCC TGA G-3') for connection. After diluting the digested product tenfold, use AFLP receptor-specific primers (5'-GAT GAG TCC TGA GTA AN-3', referred to as MseI-N) for PCR amplificatio...

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Abstract

The invention discloses Hyla tsinlingensis microsatellite sites, primers for the microsatellite sites and application of the microsatellite sites and the primers. The Hyla tsinlingensis microsatellite sites include 12 stable polymorphic microsatellite sites of which the sequences are respectively shown in SEQ ID NO:1-SEQ ID NO:12. By using the Hyla tsinlingensis microsatellite sites and through analyzing genetic typing results of 18 Hyla tsinlingensis individuals, the number of alleles is 3-12, expected heterozygosity He is 0.145-0.903, observed heterozygosity Ho is 0-0.579. All the sites show relatively high polymorphism and the requirement for individual identification is reached.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to microsatellite sites, primers and applications of Hyla qinlingensis. Background technique [0002] Qinling hyla (Hyla tsinlingensis) belongs to the genus Hyla genus Hyla. It is an endemic species in China and has a wide distribution range. It is distributed in Yuexi County and Huoshan County of Anhui Province, southern Shaanxi, southern Gansu and Chongqing. In the context of global amphibians being threatened, it is imperative to strengthen the protection of amphibians. Molecular biological information provides a certain theoretical basis for the establishment of conservation units for modified species. However, the research in this area is still blank. Contents of the invention [0003] The invention discloses 12 microsatellite sequences and 12 pairs of microsatellite primers of Hyla Qinling. These loci play an important role in the conservation genetics research, germplasm res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 张保卫康兴王慧
Owner ANHUI UNIVERSITY
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