Method for extracting DNA from shrimp shell of procambarus clarkii

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...

AI Technical Summary

Problems solved by technology

Taking shrimp tail muscle as the sample DNA source has the following disadvantages: 1. Shrimp tail muscle is prone to corruption in different degrees during the process of collection and preservation (the meat will generally experience the stiffness of the meat, the maturity of the meat, and the corruption of the meat after being separated from the body). It is a continuous process of change, pork, chicken and other livestock and poultry meat generally go through the first two stages for a long time, while shrimp and fish meat generally go through the first two stages for a short period of time, and it is easy to enter the spoilage process), and the DNA contained in it will also change with time. A certain degree of degradation occurs, especially in summer sampling; 2. The shrimp tail muscle often needs to be killed, so for some species of shrimp or shrimp that need to be kept for follow-up experiments, the tail muscle It is not feasible; 3. If the shrimp tail muscle is polluted during the sampling process, the source of pollution cannot be eliminated

However, because the shrimp shell is hard and difficult to crack, and the rich chitin in the shrimp shell is difficult to wash during the extraction process, there is no report on the successful extraction of DNA from the shrimp shell.

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