METHOD FOR PRODUCING DOUBLE-STRANDED RIBONUCLEIC ACID FROM SACCHAROMYCES CEREVISIAE YEAST CELLS Russian patent published in 2022 - IPC C12P19/34 C12N1/18 C07H21/02 C12R1/865 

FIELD: biotechnology and pharmaceutical industry.

SUBSTANCE: invention relates to the field of biotechnology and the pharmaceutical industry. A method is proposed for obtaining double-stranded ribonucleic acid (dsRNA) from yeast cells of the killer strain Saccharomyces cerevisiae RNCIM Y448. The method involves the destruction of yeast cells, centrifugation, separation of the supernatant and precipitation of total RNA. The resulting precipitate is separated and water and sodium chloride are added with stirring, settled and centrifuged. Monohydric alcohol is added to the resulting supernatant so that its concentration is 40 vol.% in the target solution, incubated and centrifuged. A buffer solution is added to the precipitate with stirring and fractionated with a solution of lithium chloride in two successive stages: at the first stage, a solution of lithium chloride is added until its concentration in the suspension reaches 2 M, at the second stage until its concentration in the suspension reaches 4 M, the resulting suspension is centrifuged to separate precipitate and repeat sequential fractionation with lithium chloride solutions in two stages. The resulting suspension is centrifuged and resuspended in a buffer solution, then deproteinization is carried out twice with chloroform in the ratio target solution: chloroform from 1:2 to 1:10 by volume and dsRNA is precipitated from the suspension with a solution of monohydric alcohol until its concentration in the target solution is 75 vol.%, the resulting precipitate is washed twice with 96% monohydric alcohol solution.

EFFECT: invention ensures achievement of high yield and purity of the target dsRNA product.

8 cl, 2 dwg, 1 tbl, 2 ex