Solution for extraction of rna

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.

1 . A solution for extracting RNA from a biological sample containing RNA and at least DNA, said solution comprising:
 (a) phenol in an amount of more than 50% by volume based on the total amount of said solution;   (b) a polyol 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.   
     
     
         2 . The solution according to  claim 1 , wherein the phenol concentration is 55 to 65% by volume based on the total amount of said solution. 
     
     
         3 . The solution according to  claim 1 , further comprising an organic solvent for separating an aqueous layer. 
     
     
         4 . The solution according to  claim 1 , wherein said biological sample is a culture liquid of cultured cells. 
     
     
         5 . The solution according to  claim 1 , wherein said biological sample is a body fluid component of an organism. 
     
     
         6 . The solution according to  claim 1 , wherein said biological sample is a blood component of an organism. 
     
     
         7 . 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 more than 50% by volume based on the total amount of said solution; 
 (b) a polyol 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.   
     
     
         8 . 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 more than 50% by volume based on the total amount of said solution; 
 (b) a polyol 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; 
   centrifuging the obtained homogenate; and   recovering an RNA-containing aqueous layer produced by the centrifugation.   
     
     
         9 . The method according to  claim 7  or  8 , wherein the phenol concentration is 55 to 65% by volume based on the total amount of said solution of (a) to (e).