METHOD FOR DNA RECOVERY FROM BONE MATERIAL Russian patent published in 2020 - IPC C12N15/10 C12P19/34 C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to molecular biology. According to a method for DNA extraction from bone material, its milling is carried out in a liquid nitrogen medium, preparation of the lysate is carried out by incubating in a thermostat followed by centrifugation, wherein the incubation is carried out in two steps, where at the first stage, a portion of ground bone material with addition of 50 mM Tris-HCl buffer, 50 mM EDTA, pH 8.0, 1 % SDS and 20 mcl of proteinase K is incubated with stirring for 8–12 hours at temperature of 56 °C, at a second step, adding buffer of 4M Gu-HCl and incubated for 10 minutes at temperature of 70 °C, followed by a step of separating the supernatant by centrifugation, separation of nucleic acids from the lysate is carried out by passing the lysate through the silicate membrane, washing is carried out by adding to the center of the silicate membrane buffer of 10 mM Tris-HCl, pH 7.5, 80 % ethanol and subsequent centrifugation, wherein washing is performed twice, and elution is carried out by adding to the center of silicate membrane buffer 10 mM Tris-HCl, 0.5 mM EDTA, pH 8.0 and incubation at room temperature for 5 minutes, followed by centrifugation of silicate membrane.

EFFECT: invention allows to enhance the quality of the obtained nucleic acid preparation and reduce the preparation time thereof.

5 cl, 1 dwg