Method for nondestructive extraction of rapana venosa genome DNA

A technology for the genome of the red snail, which is applied in the field of genomic DNA extraction of the red snail, can solve the problems of increasing the research cost and being unfavorable for the protection of the germplasm resources of the red snail, and achieves the protection of tissue integrity, low cost and simple operation. Effect

Inactive Publication Date: 2015-10-14
OCEAN UNIV OF CHINA
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

This destructive sampling method not only increases the research cost, but also is not conducive to the protection of the germplasm resources of the red snail

Method used

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  • Method for nondestructive extraction of rapana venosa genome DNA
  • Method for nondestructive extraction of rapana venosa genome DNA
  • Method for nondestructive extraction of rapana venosa genome DNA

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Embodiment 1

[0021] Select 6 red snails at random from the culture pond, adopt the method of the present invention to extract genomic DNA, the specific steps are as follows: first wipe the shell mouth and the gastropods of the red snails with a cotton ball soaked in 75% alcohol for disinfection, Put the red snail in seawater with a salinity of 50ppt for 5 minutes, then take the red snail out of the seawater with a salinity of 30ppt and put it in seawater with a salinity of 30ppt for 20 minutes. Take it out from the sea water, break off the shell of the snail, gently touch the gastropod of the snail with a sterilized small stick, stimulate the snail to secrete mucus; then use a 1.5 mL centrifuge tube to collect along the shell mouth of the snail 300-500 μL of mucus; add 400 μL of extraction buffer (6 mol / L urea, 10 mmol / L Tris-HCl, 125 mmol / L NaCl, 1% SDS, 10 mmol / L NaCl to the collected snail mucus) 2 EDTA·2H 2 O, pH 7.5), 10 μL of proteinase K, mixed well, digested at 37°C for 12 h, and ...

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Abstract

The invention discloses a method for nondestructive extraction of rapana venosa genome DNA. The method comprises: scrubbing a shell opening and a pleopod of a rapana venosa with a cotton ball soaked with 75% of alcohol for disinfection; putting the rapana venosa into seawater with the salinity being 50 ppt for 5-8 min, then putting the rapana venosa into seawater with the salinity being 29-31 ppt for 15-20 min, taking the rapana venosa out from the seawater till the pleopod stretches out of the rapana venosa, and allowing the rapana venosa to generate mucus via applying physical stimulation; collecting mucus of 300-500 [mu]L along the shell opening with a centrifuge tube of 1.5 mL; and extracting DNA from the collected rapana venosa mucus via a normal phenol-chloroform method. Experiments show that the method causes no damage to a rapana venosa individual and has characteristics, such as low cost, simple operation, and high quality of the extracted genome DNA. The method for nondestructive extraction of the rapana venosa genome DNA not only can provide a beneficial auxiliary means for rapana venosa genetic resource conservation, but also can provide powerful technical support for researches, such as molecular assistant breeding of a rapana venosa.

Description

technical field [0001] The invention belongs to the technical field of shellfish breeding, and in particular relates to a method for extracting the genome DNA of the red snail. Background technique [0002] Vein red snail ( Rapana venosa ), commonly known as conch and red snail, with an adult shell height of about 11 cm, is a large marine benthic mollusc, widely distributed in my country's Bohai Sea, Yellow Sea, East China Sea, North Korea, and the coast of Japan. Vein red snails have large feet and delicious meat. It is an important economic shellfish in my country and is deeply loved by consumers. In recent years, with the decline of the germplasm resources of the red snail in my country and the continuous expansion of market demand, the red snail has become an important breeding and development species in my country, and related work such as seed breeding has begun, and molecular biology techniques have also It is widely used in the research of genetic diversity analysis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 于红杨智鹏李琪于瑞海李浩浩
Owner OCEAN UNIV OF CHINA
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