Method of recovering nucleic acids and kit for recovering nucleic acids
View Patent ↗Disclosure is a method of separately recovering deoxyribonucleic acid and ribonucleic acid from a biological sample solution. They are separated and recovered from a single biological sample by controlling pH values, temperature, and/or concentration of cationic ions of a nucleic acid sample solution containing a caotropic agent. It is possible to easily and safely separate and recover deoxyribonucleic acid and ribonucleic acid from the single biological sample.
1. A method of recovering nucleic acids from a sample solution, said method comprising:
contacting a first sample solution comprising ribonucleic acid (RNA) and double stranded deoxyribonucleic acid (DNA) with quartz glass wool in a first binding reaction, wherein the first binding reaction results in binding of said DNA to said quartz glass wool thereby creating a second sample solution comprising the RNA, wherein during the first binding reaction the first sample solution has a pH value of 6.0 or higher but 6.9 or lower, a temperature of 50° C. or higher but 70° C. or lower and a magnesium ion concentration of 100 mM or higher but 250 mM or lower;
contacting the second sample solution with quartz glass wool in a second binding reaction, wherein the second binding reaction results in the RNA binding to said quartz glass wool in the second sample solution, wherein during the second binding reaction the second sample solution has a pH value of 5.1 or higher but 6.0 or lower, a temperature of 20° C. or higher but 50° C. or lower and a magnesium ion concentration of 100 mM or lower; and
recovering said DNA bound to the quartz glass wool in the first binding reaction, and recovering the RNA bound to the quartz glass wool in the second binding reaction.
2. The method of recovering nucleic acids according to claim 1 , wherein in the first binding reaction said DNA is bound to the quartz glass wool by contacting the first sample solution with the quartz glass wool and said DNA is eluted from the quartz glass wool after the first binding reaction; and in the second binding reaction the RNA is bound by contacting the second sample solution with the quartz glass wool from which said DNA has been eluted.
3. The method of recovering nucleic acids according to claim 1 , wherein the deoxyribonucleic acid is bound to a first quartz glass wool by contacting the first sample solution with the first quartz glass wool in the first binding reaction; and the RNA is bound to a second quartz glass wool by contacting the second sample solution with the second quartz glass wool in the second binding reaction.
4. The method of recovering nucleic acids according to claim 1 , wherein the pH value of the sample solution is lowered before the second binding reaction.
5. The method of recovering nucleic acids according to claim 1 , wherein the magnesium ion concentration of the second sample solution is lowered by diluting the second sample solution before the second binding reaction.
6. The method of recovering nucleic acids according to claim 1 , wherein the magnesium ion concentration of the second sample solution is lowered by adding a chelating agent to the second sample solution before the second binding reaction.
7. The method of recovering nucleic acids according to claim 3 , wherein a temperature is changed of at least one of the first quartz glass wool, second quartz glass wool and a sample solution.
8. The method of recovering nucleic acids according to claim 3 , wherein a temperature change is effected by controlling a temperature of a vessel portion where the first quartz glass wool and/or second quartz glass wool is disposed.
9. The method of recovering nucleic acids according to claim 3 , wherein a temperature change is effected by controlling a temperature at a portion between the first quartz glass wool and second quartz glass wool, the portion being in a vessel where the first and second quartz glass wool are disposed.