Method for extracting vibrio parahaemolyticus total RNA

Abstract

The invention provides a method for extracting vibrio parahaemolyticus total RNA, which is characterized by comprising the following steps of: (1) putting 1ml of vibrio parahaemolyticus liquid in a centrifuge tube to be centrifuged; (2) removing supernatant, quickly freezing thallus with liquid nitrogen, grinding into powder, transferring the powder into the centrifuge tube, adding Trizol, shaking for 1min, standing on ice for 3min, and repeatedly shaking for three times; (3) adding chloroform / isoamylol, and centrifuging; (4) transferring 500ul of supernatant into the centrifuge tube, adding 500ul of isopropanol, standing to fully precipitate and centrifuging; (5) carefully removing supernatant, washing with 70 percent of ethanol, and centrifuging; and (6) carefully absorbing supernatant as far as possible, inversely standing on a ultra-clean table to dry alcohol, adding DEPC water for dissolving, and storing under the temperature of 80 DEG C below zero. Since adopting the technical scheme, the invention has the advantages and the beneficial effects that the method can extract total RNA with high concentration, high purity and good integrity in the short three hours, and the used reagent is simple and has low cost.

Description

technical field [0001] The invention relates to a method for extracting total RNA of vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus (Vibrio parahaemolyticus) is a Gram-negative polymorphic bacillus or slightly curved Vibrio, which widely exists in seawater and grows well in a medium containing 3% to 3.5% salt concentration, so it is also called pathogenic Halobacteria. Vibrio parahaemolyticus is a common pathogenic bacteria in aquatic products, pickled vegetables, cooked meat, poultry meat and poultry eggs. or undercooked food can cause food poisoning, so it is of great significance to carry out biological research on Vibrio parahaemolyticus. Isolation and extraction of high-quality RNA is the basis for subsequent molecular biology research such as PCR, Real-time PCR, gene cloning, and cDNA library establishment. Bacterial RNA has the characteristics of less content, short half-life, and easy degradation, so it is more difficult to extract ...

AI Technical Summary

Problems solved by technology

Bacterial RNA has the characteristics of less content, short half-life, and easy degradation, so it is more difficult to extract high-quality RNA than eukaryotes such as plants and animals.

At present, there is no report on the extraction of RNA from Vibrio parahaemolyticus at home and abroad