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Duplex PCR microsatellite marker for identifying acipenser sinensis genetic relationship and identification method thereof

A technology of microsatellite marker and kinship, applied in the field of double PCR microsatellite marker, to achieve the effect of reducing financial resources and manpower and huge promotion value

Active Publication Date: 2018-10-09
CHINESE STURGEON RES INST CHINA THREE GOR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the microsatellite marker multiplex PCR of Chinese sturgeon at home and abroad

Method used

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  • Duplex PCR microsatellite marker for identifying acipenser sinensis genetic relationship and identification method thereof
  • Duplex PCR microsatellite marker for identifying acipenser sinensis genetic relationship and identification method thereof
  • Duplex PCR microsatellite marker for identifying acipenser sinensis genetic relationship and identification method thereof

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Experimental program
Comparison scheme
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Embodiment 1

[0019] Forty-eight Chinese sturgeons were collected from the Chinese Sturgeon Research Institute of China Three Gorges Corporation, and the fin rays of Chinese sturgeons were collected to extract the genomic DNA of Chinese sturgeons using the traditional phenol-chloroform extraction method.

[0020] Take 0.1g of caudal fin from each individual, put it in a 1.5ml centrifuge tube, cut it into pieces, add 450μL STE extraction buffer (10mmol / L Tris-HCl, pH8.0; 1mmol / L EDTA, pH8.0), 35μL SDS (10 %), 15 μL proteinase K (0.2%).

[0021] Place the centrifuge tube in a water bath at 55°C for 1 hour until clear and transparent.

[0022] Add 700ul Tris saturated phenol into the centrifuge tube, mix on a shaker for 30 minutes, centrifuge at 12,000 rpm for 10 minutes at 4°C, transfer the supernatant to another clean eppendorf tube (note the 1mL tube with a tip cut flat) Aspirate the supernatant in order not to confuse the lower sediment).

[0023] Add an equal volume of phenol-like alcoh...

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Abstract

The invention discloses a duplex PCR microsatellite marker for identifying the acipenser sinensis genetic relationship and an identification method thereof, and belongs to the field of animal molecular genetics. The invention provides the duplex PCR microsatellite marker of the acipenser sinensis and the method for identifying the genetic relationship between acipenser sinensis individuals by using the marker. By using the provided microsatellite marker, the genetic relationship between the individuals can be fast identified; the inbreeding between the individuals can be avoided. The microsatellite marker amplification result provided by the invention has high polymorphism and stability; in addition, the money and the labor consumed by the duplex PCR microsatellite marker provided by the invention are halves of those of the conventional PCR reaction; great practical values are realized.

Description

technical field [0001] The invention relates to a double PCR microsatellite marker for identifying the genetic relationship of Chinese sturgeon and an application thereof, belonging to the field of animal molecular genetics. Background technique [0002] The Chinese sturgeon is the fish with the lowest distribution latitude and the largest body among more than 20 species of sturgeons in the world. It is a species unique to my country. In the 1970s, the breeding population in the Yangtze River could reach more than 10,000. Due to environmental pollution, overfishing, habitat loss and many other factors, the Chinese sturgeon is on the verge of extinction. The protection of endangered species not only protects the individuals of endangered species, but also understands and protects the genes of endangered species, and then formulates good breeding policies to prevent inbreeding. However, until now, the genetic research on Chinese sturgeon is very limited, and it is urgent to s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2537/143C12Q2525/151
Inventor 胡亚成杜合军杨菁刘雪清肖衎
Owner CHINESE STURGEON RES INST CHINA THREE GOR