Solution for extraction of RNA
AssigneeToray Industries, Inc.
InventorsMakiko Yoshimoto, Hideo Akiyama, Hitoshi Nobumasa
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Abstract
A solution for extracting substantially pure RNA from a biological sample is disclosed. The solution for extracting RNA from a biological sample containing RNA and at least DNA comprises: (a) phenol in an amount of more than 50% by volume based on the total amount of the solution; (b) a polyol in an amount of 3 to 10% by volume based on the total amount of the solution; (c) a guanidinium salt at a concentration of 0.5 to 2.0 M based on the total amount of the solution; (d) a thiocyanate at a concentration of 0.1 to 0.5 M based on the total amount of the solution; and (e) a buffer for maintaining the pH of the solution at 4 to 6.
Claims9
A method for extracting RNA from a biological sample containing RNA and at least DNA, said method comprising the steps of:
- homogenizing said biological sample together with a solution comprising:
- (a) phenol in an amount of not less than 53% by volume based on the total amount of said solution, wherein the amount of phenol does not exceed a percentage that excludes any of components (b)-(e),
- (b) an aliphatic alcohol having a plurality of hydroxyl groups in an amount of 3 to 10% by volume based on the total amount of said solution;
- (c) a guanidinium salt at a concentration of 0.5 to 2.0 M based on the total amount of said solution;
- (d) a thiocyanate at a concentration of 0.1 to 0.5 M based on the total amount of said solution;
- and
- (e) a buffer for maintaining the pH of said solution at 4 to 6;
- mixing the obtained homogenate with an organic solvent for separation of an aqueous layer;
- centrifuging the obtained mixture;
- and
- recovering an RNA-containing aqueous layer produced by the centrifugation.
A method for extracting RNA from a biological sample containing RNA and at least DNA, said method comprising the steps of:
- homogenizing said biological sample together with a solution comprising:
- (a) phenol in an amount of not less than 53% by volume based on the total amount of said solution, wherein the amount of phenol does not exceed a percentage that excludes any of components (b)-(e);
- (b) an aliphatic alcohol having a plurality of hydroxyl groups in an amount of 3 to 10% by volume based on the total amount of said solution;
- (c) a guanidinium salt at a concentration of 0.5 to 2.0 M based on the total amount of said solution;
- (d) a thiocyanate at a concentration of 0.1 to 0.5 M based on the total amount of said solution;
- (e) a buffer for maintaining the pH of said solution at 4 to 6;
- and
- (f) an organic solvent for separation of an aqueous layer, wherein the organic solvent is in addition to the phenol;
- centrifuging the obtained homogenate;
- and
- recovering an RNA-containing aqueous layer produced by the centrifugation.
Dependencies are inferred from the claim text.