Extraction method of total RNA of intermuscular bone of megalobrama amblycephala

Abstract

The invention discloses a method for extracting total RNA of the intermuscular bone of megalobrama amblycephala, which comprises the following steps: 1) with megalobrama amblycephala as the material, fetching the intermuscular bone thereof and putting into a sterilized and pre-cooled mortar, and grinding until the intermuscular bone is ground into powder; transferring the powder into a centrifugal tube; adding Trizol and shaking uniformly; and placing at room temperature; 2) centrifuging at 4 DEG C, fetching the supernate, adding chloroform the volume of which is 1 / 5 that of the supernate, shaking so that the mixture is sufficiently emulsified into a uniform white emulsion without layering, and standing at room temperature; 3) centrifuging at 4 DEG C, fetching the supernate and repeatedly extracting with chloroform; 4) fetching the supernate, adding isometric isopropyl alcohol, sodium citrate and sodium chloride and mixing uniformly, and standing at -20 DEG C; 5) after the standing, centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, adding ethanol prepared by DEPC water, shaking and repeatedly washing and precipitating; and 6) centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, naturally airing, and adding DEPC water to dissolve RNA to obtain the total RNA solution. The process is simple, the cost is low, the obtained total RNA has high concentration, high purity and high integrity, and further molecular biology study is met.

Description

technical field [0001] The invention relates to a method for extracting total RNA from fish bones, in particular to a method suitable for extracting total RNA from intermuscular bones of bream. This method is also applicable to the extraction of total RNA from the intermuscular bone of other fishes, and the extracted total RNA can be used for cDNA synthesis and gene expression research in further molecular biology. Background technique [0002] Fish meat is more and more popular because of its high nutritional content, low fat content, easy digestion, and is suitable for the general population. Dory ( Megalobrama amblycephala ) belongs to Cyprinidae ( Cypriniformes ), Cyprinidae ( Cyprinidae ), Subfamily Cichlidae ( Culterinae ), Bream ( Megalobrama ). It is one of the important herbivorous economic fishes unique to my country. Because of its wide feeding habits, low cost, fast growth, high survival rate, easy to catch, and has the advantages of delicious taste, sm...

AI Technical Summary

Problems solved by technology

The method of extracting total RNA from the intermuscular bone which is small in size, easy to degrade and easily contaminated by muscle protein has not yet appeared.

The amount of total RNA extracted from intermuscular bone by traditional methods is small and highly polluted, which cannot support further molecular experiments

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