Method and test kit for rapid isolation of nucleic acids using rough surfaces
The invention relates to a method for isolation of nucleic acids from aqueous samples containing nucleic acids in free form or liberated by lysis. Before or after the polarity of the aqueous solution is lowered, the sample is brought into contact with a solid phase that has a rough or structured surface, whereupon the nucleic acids precipitate on the solid phase and then, together with the solid phase, are removed from this aqueous solution. The rough or structured surface is preferably a non-smooth metal, plastic or rubber surface. Subject matter of the invention is also a test kit and a corresponding instrument for isolation of nucleic acids.
1. A method for isolation of a nucleic acid from an aqueous sample that comprises a nucleic acid, the method comprising:
contacting an aqueous solution that comprises a nucleic acid in free form or liberated by lysis with a solid phase that has a rough or structured surface;
lowering the polarity of the aqueous solution before or after said contacting;
whereupon said contacting, precipitating the nucleic acid on the solid phase; and, thereafter,
removing the precipitated nucleic acid together with the solid phase from the aqueous solution.
2. The method according to claim 1 , wherein
the nucleic acid comprises a short-chain and a long-chain nucleic acid, which bind simultaneously on the solid phase.
3. The method according to claim 1 , wherein
said lowering is carried out with an organic solvent, thereby
lowering the polarity of the aqueous solution.
4. The method according to claim 3 , wherein
the organic solvent is present, during said lowering, in an amount of from 5 vol % and 90 vol %.
5. The method according to claim 1 , wherein
the aqueous solution further comprises at least one of:
a) a salt;
b) at least one detergent;
c) an amino alcohol or a substance for adjustment of the pH.
6. The method according to claim 5 , wherein
a) the salt is present in a concentration of from 1 mM and 5 M,
b) the detergent is present in a concentration of from 0.1 vol % and 30 vol %,
c) wherein when an amino alcohol or substance is present, TRIS is present in a concentration of from 1 mM and 2 M.
7. The method according to claim 1 , wherein
the aqueous solution further comprises an additional additive.
8. The method according to claim 1 , wherein
the rough or structured surface is a non-smooth metal, plastic or rubber surface or magnetic particles.
9. The method according to claim 8 , wherein
the non-smooth surface is produced by 3D printing or by roughening a plastic.
10. The method according to claim 1 , wherein
the solid phase comprises a rough composite material.
11. The method according to claim 1 , wherein
the solid phase comprises a roughened pipette tip.
12. The method according to claim 1 , wherein
the solid phase with a structure surface is the thread of a screw, a metal sponge or granules, a twisted material or a 2- or 3-dimensional network structure.
13. The method according to claim 1 , further comprising:
moving the solid phase in the aqueous solution.
14. A test kit for isolation of a nucleic acid from an aqueous sample comprising a nucleic acid, the test kit comprising:
at least one substance for lowering the polarity of the aqueous solution,
at least one solid phase with a rough or structured surface for binding the nucleic acid, and
an elution buffer.
15. The method according to claim 1 , wherein said solid phase has a structured surface which has grooves.