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Hemibarbus maculates microsatellite sites and primers

A technology of microsatellite sites and fish bones is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Achieve high polymorphism and stability

Inactive Publication Date: 2011-01-26
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of physical markers to identify larvae has major limitations, such as the short duration of markers, high cost, and heavy workload. The larvae can only be marked when they reach the size of the marker, and environmental errors cannot be completely eliminated.

Method used

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  • Hemibarbus maculates microsatellite sites and primers
  • Hemibarbus maculates microsatellite sites and primers
  • Hemibarbus maculates microsatellite sites and primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] 1. Screening of microsatellite loci

[0022] 1. Genomic DNA extraction, digestion and recovery:

[0023] Use phenol extraction to extract genomic DNA from flower fish bones: each individual weighs 100mg of muscle, washes it twice with sterile water, cuts it into pieces and adds 10mM Tris-Hcl pH 8.0, 20mM EDTA pH 8.0, 10mM NaCl, 1% SDS, 600 μl of lysate of 100 μg / ml proteinase K, after digesting the muscle at 55 ° C, centrifuge at 12000 rpm for 10 min, take the supernatant into a new 1.5 ml centrifuge tube, add phenol, chloroform and isoamyl alcohol at a ratio of 25:24:1 And extract twice, the ratio is 24:1 chloroform and isoamyl alcohol After extraction once, the water phase is transferred to a new tube, add -20 ℃ absolute ethanol, centrifuge at 12,000rpm for 10min, wash the precipitate twice with 75% ethanol , dried at room temperature and dissolved in 100 μl 0.1×TE.

[0024] 2. Digestion, recovery and linker connection:

[0025] Take 100 μg of genomic DNA, and part...

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Abstract

The invention relates to hemibarbus maculates microsatellite sites and primers. The invention has the characterized in that the nucleotide sequence of the microsatellite sites is SEQ ID NO:1-10, and the sequence of the microsatellite primers is SEQ ID NO:11-30. In the invention, 10 microsatellite sites are screened out form the DNA of a hemibarbus maculates genome, and specific primers are designed according to the flanking sequence of the sites on both ends of each microsatellite site. Amplification results show that the acquired primers have high polymorphism and stability and can be applied to the fields of population genetic diversity detection, individual identification and molecular-assisted breeding of hemibarbus maculates.

Description

technical field [0001] The invention belongs to the technical field of DNA markers in molecular biology, and in particular relates to microsatellite sites and primers of flower fish bones. Background technique [0002] Hemibarbus maculates, which belongs to Cyprinidae, Scorpioninae, and Hemibarbus genus, is widely distributed in rivers and lakes in my country, and is a common small and medium-sized wild economic fish in natural water bodies. But in the past ten years, due to overfishing, dam building and sand dredging, and water pollution, the wild resources of flower fish bones have been seriously damaged. In order to meet the needs of the market, large-scale artificial breeding has been carried out on the flower fish bones, but the current seedlings have not been artificially selected, and there are phenomena such as low growth rate and poor disease resistance. It has become one of the important issues to be solved urgently for the healthy and sustainable development of my...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 朱俊杰练青平张爱菊原居林刘金殿
Owner ZHEJIANG INST OF FRESH WATER FISHERIES