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New solutions to provoke binding of nucleic acid to a solid support

IP.com Disclosure Number: IPCOM000251085D
Publication Date: 2017-Oct-10

Publishing Venue

The IP.com Prior Art Database

Abstract

Common methods for the isolation of nucleic acids from nucleic acid containing samples such as cells, tissues, or cell free liquid comprise: - contacting the sample with a chaotropic solution, - dissolution of the sample therein, - addition of an organic solvent to the mixture, - contacting the mixture with a solid support and binding of nucleic acid to the solid support. - Subsequently, the solid support is washed to remove contaminants and finally nucleic acid can be eluted from the solid support. Most commonly used organic solvents to provoke binding of nucleic acid to a solid support out of chaotropic lysates are ethanol and isopropanol. Further organic solvents, like higher alcohols, ketones, and lactones have been described to be useful for such an application. Here we describe the use of Benzyl alcohol, Diethyl malonate, Dimethyl adipate, Dimethyl glutarate, Dimethyl succinate, Dipropylen glycol, Ethylene glycol acetate, Solketal, Triacetin, Triethyl citrate, and Triethylene glycol or mixtures thereof as alternative organic solvents, that provoke nucleic acid binding to a solid support. These solvents are especially advantageous in isolation processes starting with very tiny amounts of sample material.

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New solutions to provoke binding of nucleic acid to a solid support

Abstract - Introduction

Common methods for the isolation of nucleic acids from nucleic acid containing samples such as cells, tissues, or cell free liquid comprise:

- contacting the sample with a chaotropic solution, - dissolution of the sample therein, - addition of an organic solvent to the mixture, - contacting the mixture with a solid support and binding of nucleic acid to the solid

support. - Subsequently, the solid support is washed to remove contaminants and finally

nucleic acid can be eluted from the solid support.

Most commonly used organic solvents to provoke binding of nucleic acid to a solid support out of chaotropic lysates are ethanol and isopropanol. Further organic solvents, like higher alcohols, ketones, and lactones have been described to be useful for such an application.

Here we describe the use of Benzyl alcohol, Diethyl malonate, Dimethyl adipate, Dimethyl glutarate, Dimethyl succinate, Dipropylene glycol, Ethylene glycol acetate, Solketal, Triacetin, Triethyl citrate, and Triethylene glycol or mixtures thereof as alternative organic solvents, that provoke nucleic acid binding to a solid support. These solvents are especially advantageous in isolation processes starting with very tiny amounts of sample material.

State of the art – background

The use of alcohol (e.g. ethanol, isopropanol) as a binding solution for the isolation of RNA in combination with silica and chaotropic agents is state of the art, published numerously and is used in e.g. the NucleoSpin RNA kits (MN), RNeasy kits (Qiagen), and many other kits.

EP 1502951 A1, EP 1526176 A2, EP 1626085 A1 (Agilent) discloses methanol, ethanol, isopropanol and polyethylene glycol as organic binding enhancer in a chaotropic solution to bind nucleic acids to a filtration column.

EP 2041281 (AJ Innuscreen) discloses the use of alcohols with 1 – 5 carbon atoms in combination with salt to bind RNA to a solid support.

EP 0743950 B1 and EP 1146049 (Qiagen) discloses lower aliphatic alcohols having from 1 – 5 carbon atoms in combination with chaotropic and mineral supports for the isolation of RNA.

EP 1524317 A1 (Roche) discloses the use of acetone, acetylacetone, acetonitrile, dimethylsulfoxide, diethylketone, methylethylketone, methylpropylketone, isobutylmethylketone, gamma-butyrolactone, gamma-valerolactone, propylene

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carbonate, and N-methyl-2-pyrrolidone for isolation of nucleic acid by adsorbing nucleic acid to a mineral substrate from a composition containing an aqueous buffer, salts in high concentration and one of the disclosed organic compounds.

EP 2137309 (Stratagene) discloses the use of sulfolane for the isolation of nucleic acid by applying a sample containing a nucleic acid in the presence of an mixture of salts and sulfolane with a mineral substrate that adsorbs at least one nucleic acid.

EP 2217703 A1 (GE Healthcare) discloses the use of Acetone, Acetonitrile, Tetrahydrofuran (THF), Met...