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Rattus losea microsatellite DNA marker and amplification primer thereof and a detection method and application thereof

A technology of DNA labeling and amplification primers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., to achieve good reproducible results

Active Publication Date: 2018-07-20
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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Problems solved by technology

There are currently no domestic research reports on the molecular ecology of yellow-haired mice, and there are relatively few foreign studies in this area. In order to further understand the information on the population structure and genetic diversity of yellow-haired mice, it is necessary to develop new molecular genetic markers

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  • Rattus losea microsatellite DNA marker and amplification primer thereof and a detection method and application thereof
  • Rattus losea microsatellite DNA marker and amplification primer thereof and a detection method and application thereof
  • Rattus losea microsatellite DNA marker and amplification primer thereof and a detection method and application thereof

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Embodiment 1

[0065] 1. Experimental steps

[0066] 1.1. DNA extraction from yellow-haired mice

[0067] About 50 mg of yellow-haired mouse muscle was cut for DNA extraction, and the extraction method used a standard chloroform-isoamyl alcohol-alcohol extraction procedure. Use agarose gel electrophoresis and ultraviolet spectrophotometer to detect the purity and concentration of DNA, and dilute it into a 30ng / μL working solution, and store it at 4°C for use. DNA concentration was detected by 1% agarose gel electrophoresis (see figure 1 ).

[0068] 1.2. Selection of microsatellite sites

[0069] Referring to the microsatellite loci of the closely related species of yellow-haired rats, the SSR loci were searched using misa software. On both sides of the SSR site, 60 pairs of primers were designed using Primer 3 software.

[0070] 1.3. Primer screening

[0071] The preliminary screening of 60 pairs of primers was carried out with templates of better quality. The amplification system and ...

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Abstract

The invention discloses a rattus losea microsatellite DNA marker and an amplification primer thereof and a detection method and application thereof. Through the construction of rattus losea microsatellite (CT) n, (AG) n, (TG) n and (AC) n enriched libraries, screening and sequencing of positive clones containing microsatellite sequences and the cloning containing a microsatellite repeated sequence, eight microsatellite markers chrom1-26, chrom2-2, chrom15-6, chrom16-6, chrom16-12, chrom20-9, chrom20-37 and chrom-16 rich in polymorphism are obtained, and nucleotide sequences thereof are respectively as shown in SEQ ID No. 1-8. The rattus losea microsatellite DNA marker can be applied to researches of study of genetic diversity and the identification of parental relationship of different populations of rattus losea, is good in repeatability and is a reliable and effective molecular marker.

Description

Technical field: [0001] The invention belongs to the technical field of molecular markers, and in particular relates to yellow-haired mouse microsatellite DNA markers, amplification primers, detection methods and applications thereof. Background technique: [0002] Microsatellite DNA widely exists in prokaryotic and eukaryotic genomes, common two-, three- and four-nucleotide repeat sequences, accounting for 50% of eukaryotic genomes, arranged by tandem 2-6bp simple sequence repeats (such as: AC , CCA or GATA, SSR). The most common are dinucleotide repeats, such as (AC / TG)n and (AG / TC)n. The microsatellite DNA at each specific locus consists of two parts: the central core region and the peripheral flanking regions. There may be different individuals in an organism with the same flanking regions but different numbers of tandem repeat units; it is also possible to have the same number of repeat units but different sizes of flanking regions, or both. DNA RFLP studies of non-m...

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 秦姣刘全生张春兰
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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