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Method for extracting sinonovacula constricta DNA in vivo

A technique for constricting razor clams and living bodies, which is applied in the field of extracting biological DNA, can solve the problems of detecting growth traits, the effect of germplasm utilization, and the like, and achieve the effect of improving breeding efficiency

Inactive Publication Date: 2014-02-26
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This has a great impact on the detection of its growth traits and the subsequent use of germplasm

Method used

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  • Method for extracting sinonovacula constricta DNA in vivo
  • Method for extracting sinonovacula constricta DNA in vivo
  • Method for extracting sinonovacula constricta DNA in vivo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] In order to verify that the DNA extracted from the mantle and the outlet tube of the same individual is the same, this Example 1 uses a pair of common primers to perform PCR detection on the DNA extracted from the mantle and outlet tube of 5 constriction razor clams.

[0027] 1. Obtaining the mantle and outlet pipe of Sinonovacula constricta

[0028] The Ministry of Education Key Laboratory of Aquatic Germplasm Resources Discovery and Utilization was jointly established in Shanghai Ocean University, selected Ningbo Changjie constriction as the experimental material, and temporarily raised 20 constriction in a seawater culture tank with a water temperature of 25℃ and a specific gravity of 1.015. 3 Five days later, 5 constrictions were randomly selected. The outlet pipe of Sinonovacula constricta is labeled s1-5; the mantle of Sinonovacula constricta is labeled m1-5.

[0029] 2. DNA extraction

[0030] Add 250μl of DNA extraction solution to the tissue collected in the water out...

Embodiment 2

[0041] In order to confirm that the removal of the outlet pipe of the constriction does not affect the survival of the constriction, we performed the operation of cutting out the water pipes of 20 constrictions, and temporarily raising the treated individuals.

[0042] 1. Put the constrictor snails from the tissue to be taken into a seawater culture tank with a water temperature of 25°C for 3 days, and stick a numbered mark on the shell of the constrictor snails. When DNA needs to be extracted, place the constriction of the tissue to be taken on ice, anesthetize the constriction for 3-5 minutes, use scissors to cut out the water pipe of the constriction 0.5cm, and put the treated constriction back into the culture barrel.

[0043] 2. Inflate the culture barrel and feed Chrysophyllum and Chaetoceros daily for 2 weeks. During this period, the survival rate of Sinonovacula constricta is recorded, see image 3 .

[0044] 3. The experimental results show that cutting the outlet pipe of S...

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Abstract

A disclosed method for extracting sinonovacula constricta DNA in vivo comprises the following steps: 1) isolating of sinonovacula constricta water outlet vessel tissue; 2) extraction of DNA in water-outlet vessel tissue; and 3) detection of extracted DNA. The method helps to extract DNA from sinonovacula constricta water-outlet vessel tissue being regenerative and avoid killing of sinonovacula constricta or influence on growth of sinonovacula constricta. The genotype of the sinonovacula constricta DNA extracted by employing the method can be detected, so that the method has important application value in sinonovacula constricta scientific research and breeding. In family cultivation, parent is extremely precious, and the harm to the parent influences the investigation on production performance and germplasm utilization. The method provided by the invention can help to detect the genotype of family parent, so that the production performance of filial generation can be predicted, the family is selected and the sinonovacula constricta breeding efficiency is improved.

Description

Technical field [0001] The invention relates to a method for extracting biological DNA, in particular to a method for extracting DNA from Sinonovacula constricta from a living body. Background technique [0002] Sinonovacula constricta Lamarck (Sinonovacula constricta Lamarck) is an important tidal flat shellfish in my country, and it is one of the four aquaculture shellfish. According to statistics, the production of shellfish in my country accounts for 82% of the total production of mariculture. The annual output of tidal flat shellfish is nearly 2 million tons, of which the output of Sinonovacula constricta reaches 700,000 tons, which accounts for about 30% of my country's total tidal flat shellfish production. Although the culture of Sinonovacula constricta has a history of more than 800 years, the seedlings are all from wild species. Cultivating high-quality varieties with fast growth and good economic characteristics is an important way to ensure the sustainable developmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 李家乐王劦牛东红沈和定李多王泽
Owner SHANGHAI OCEAN UNIV
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