Method for extracting DNA from shrimp shell of procambarus clarkii

A technology of Procambarus clarkii and shrimp shells, which is applied in the field of extracting DNA from the shells of Procambarus clarkii, can solve the problems of corruption, difficult cracking, and difficult cleaning of chitin, and achieve the effect of easy operation and pollution prevention

A technology of Procambarus clarkii and shrimp shells, which is applied in the field of extracting DNA from the shells of Procambarus clarkii, can solve the problems of corruption, difficult cracking, and difficult cleaning of chitin, and achieve the effect of easy operation and pollution prevention

CN102477423AInactive Publication Date: 2012-05-30HUAZHONG AGRI UNIV
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  • Method for extracting DNA from shrimp shell of procambarus clarkii
  • Method for extracting DNA from shrimp shell of procambarus clarkii
  • Method for extracting DNA from shrimp shell of procambarus clarkii

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Example 1 Extraction of Procambarus clarkii Shrimp Shell DNA

[0029] Buy fresh Procambarus clarkii from the market, take part of the shrimp shells, and soak them in absolute ethanol for storage (the storage time is one month)

[0030] 1) Reagent preparation:

[0031] A) Tissue cell lysate: 100mM Tris-HCl, pH 8.0; 50mM EDTA, pH 8.0; 1% sodium dodecyl sulfate (SDS);

[0032] B) 0.5M EDTA: 23.3g disodium ethylenediamine tetraacetate dihydrate (EDTA-Na·2H 2 O) Dissolve in 100mL double-distilled water, stir vigorously on a magnetic stirrer, add NaOH to adjust the pH value to 8.0 while stirring, then sterilize by high pressure steam (121°C, sterilize for 20min), store at room temperature (note: dihydrate B Disodium diaminetetraacetic acid needs to add NaOH to adjust the pH value of the solution to 8.0 before dissolving).

[0033] C) 1M dithiothreitol (DTT): Add 32.4mL water to the original 5g bottle of dithiothreitol, divide into small portions and store at -20°C;

[003...

Embodiment 2

[0040] According to the step of embodiment 1, wherein the shrimp shell of Procambarus clarkii is the material preserved for 3 months with absolute ethanol.

Embodiment 3

[0042] According to the step of embodiment 1, wherein the shrimp shell of Procambarus clarkii is the material preserved as 6 months with absolute ethanol.

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Abstract

The invention belongs to the technical field of aquatic animal molecular biology DNA sample preparation, and concretely relates to a method for extracting DNA from a shrimp shell of procambarus clarkia. The method comprises the following steps: cutting the shrimp shell of the procambarus clarkii immersed in an ethanol solution, adding a proper amount of a tissue lysate, EDTA, dithiothreitol, protease K and the like in a centrifuge tube, carrying out processes of water-bath cracking, ammonium acetate extraction and the like to obtain the total DNA of procambarus clarkia. The method of the invention has the advantages that the operation is simple and convenient, the used reagent enables small toxicity, less injury on the experimental animals when sampling is carried out, the samples are easy to be preserved for long-term, the degradation degree of the extracted DNA is lower than the degradation degree of the extracted DNA from the shrimp tail muscle. The prepared DNA can be used for diversity analysis of procambarus clarkii population genetics and the relative molecular biology application.

Description

technical field [0001] The invention belongs to the technical field of DNA sample preparation of aquatic animal molecular biology, and in particular relates to a method for extracting DNA from the shell of Procambarus clarkii. The DNA prepared by the invention can be used for the genetic diversity analysis of the Procambarus clarkii population and related molecular biology applications. Background technique [0002] Procambarus clarkii (Procambarus clarkii), belonging to Crustacea, Decapoda, Crawfish family, Procambarus genus, is native to northern Mexico and southern United States, and has become one of the world's famous invasive species. Due to its large size and strong adaptability, it can endure up to 4 months of dry season and has a high reproductive rate. It can form a single-dominant population, suppress, crowd out or destroy local species, endanger the survival of local species, and affect invasive species. diversity. However, due to its high economic value, Proca...

Claims

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Application Information

Patent Timeline
30 May 2012
Publication
CN102477423A
IPC
C12N15/10; C12Q1/68
Inventors
李艳和; 王卫民