Efficient extraction method for river crab genome DNA after long-term storage in alcohol

Abstract

The invention discloses a method for extracting the river crab genome DNA after long-term storage in alcohol, which comprises the following steps: (1) tissue fetching: clipping a little second pereiopod muscle of Eriocheir sinensis stored in alcohol, and drying the alcohol; and (2) extraction of genome DNA: preparing a lysis buffer, performing water-bath digestion and centrifuging; adding a saturated NaCl solution and chloroform and centrifuging; adding chloroform and centrifuging; adding isopropyl alcohol, and centrifuging to precipitate DNA; washing, centrifuging and drying; and adding deionized water or TE solution and storing. The method disclosed by the invention solves the difficulty in the DNA extraction of Eriocheir sinensis after long-term storage in alcohol, and conveniently and effectively extracts the river crab DNA; the extracted DNA has enough amount and high quality and is barely polluted by proteins and can be applied to subsequent multiple kinds of molecular biological analysis so as to provide a technological means for the genetic diversity research, germplasm protection and resource utilization of the river crabs; the method is of important scientific significance and practical value.

Description

technical field [0001] The invention relates to a simple, rapid and high-quality method for extracting genome DNA of crabs preserved in alcohol. Background technique [0002] River crab is the general term for freshwater mitten crabs in my country, including Chinese mitten crabs ( Eriocheir sinensis ), Japanese mitten crab ( Eriocheir japanicus ) and Hepu mitten crab ( Eriocheir hepunensis ). They all belong to Arthropoda, Crustacca, Decapo, Grapsidae, mitten crab ( Eriocheir ). Eriocheir sinensis is a unique aquatic economic animal in my country. It is widely distributed in the Liaohe River, Yangtze River, Oujiang River, Minjiang River and other water systems. In addition, it is also widely distributed in Europe and other places. Japanese mitten crabs are mainly distributed in mainland Japan, the Korean Peninsula, and Vladivostok in Russia. Hepu mitten crabs are distributed in the Nanliujiang water system (such as Hepu County) and the Pearl River water system in Gu...

AI Technical Summary

Problems solved by technology

Compared with other aquatic organisms preserved with alcohol, river crabs belong to the particularity of lower crustaceans. The carapace has a certain protective effect on muscle tissue, which makes the process of alcohol infiltrating into muscle tissue relatively slow, so that muscle tissue can be recovered in a short period of time. There is a certain degree of corruption without preservation, which affects the integrity of DNA preservation, and finally makes subsequent DNA extraction difficult

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