NUCLEIC ACID PURIFICATION METHOD
A method for purifying polynucleic acids is disclosed, comprising adsorbing the polynucleic acids to a filter, washing the filter with a water-immiscible solution, and eluting the polynucleic acids from the filter. The method can be used to prepare purified RNA and/or DNA.
1 . A method of purifying polynucleic acids, the method comprising:
(a) absorbing the polynucleic acids to a filter fitted in a container;
(b) washing the filter with a first immiscible fluid wash solution; and
(c) eluting the polynucleic acids from the filter using an aqueous-based elution solution;
wherein a solution of purified polynucleic acids is obtained.
2 . The method of claim 1 , further comprising:
after step (a) and before step (b), washing the filter with an alcohol-based wash solution.
3 . The method of claim 1 , further comprising:
after step (a) and before step (b), washing the filter with a second immiscible fluid wash solution and then washing the filter with an alcohol-based wash solution.
4 . The method of any of claim 1 , 2 , or 3 , wherein the polynucleic acids are DNA and/or RNA.
5 . The method of any of claim 1 , 2 , or 3 , further comprising:
in step (c), eluting the polynucleic acids from the filter using (i) an aqueous-based elution solution and (ii) an immiscible fluid push solution.
6 . The method of claim 1 , wherein the filter is a glass microfiber filter.
7 . The method of claim 2 , wherein the first immiscible fluid wash solution comprises a fluid that (i) when contacted with water forms a substantially separate phase from the water; and (ii) has a specific gravity less than that of water.
8 . The method of claim 7 , wherein the specific gravity of the first immiscible fluid wash solution is less than that of the alcohol-based wash solution used in the method.
9 . The method of claim 2 , wherein the alcohol-based wash solution comprises ethanol and water.
10 . The method of claim 9 , wherein the alcohol-based wash solution further comprises a salt and/or a buffer.
11 . The method of claim 9 , wherein the alcohol-based wash solution further comprises a hydrophilic polymer.
12 . The method of claim 11 , wherein the hydrophilic polymer is a polyethylene glycol polymer.
13 . The method of claim 1 , wherein the aqueous-based elution solution comprises a buffer.
14 . The method of claim 1 , wherein the aqueous-based elution solution comprises DNase/RNase-free water.
15 . A reagent kit comprising an immiscible fluid wash solution and an elution solution.
16 . The reagent kit according to claim 15 , further comprising an alcohol-based wash solution.