초록 ▼

Compositions and methods for isolating nucleic acids from biological tissues and cells and for tissue/cell solubilization for other molecular biological uses, wherein the compositions comprise, in part, novel combinations of chaotropic agents and aromatic alcohols which act synergistically to effect

Compositions and methods for isolating nucleic acids from biological tissues and cells and for tissue/cell solubilization for other molecular biological uses, wherein the compositions comprise, in part, novel combinations of chaotropic agents and aromatic alcohols which act synergistically to effect better tissue/protein solubilization. The inventive compositions further include aprotic solvents for deactivation of ribonucleases and denaturization of proteins, as well as detergents for enhancing cell lysis and nucleoprotein dissociation. The inventive methods also comprise the use of a centrifuge, a solid-support matrix, and a microporous membrane for final isolation of the precipitated nucleic acids, resulting in high yield and purity of the precipitated nucleic acid.

대표청구항 ▼

A method for isolating DNA from biological tissues and cell samples, comprising the steps of: (A) solubilizing said biological sample with a sufficient amount of a single composition to denature and solubilize said sample to form a solubilized sample, said solubilized sample comprising RNA, DNA and

A method for isolating DNA from biological tissues and cell samples, comprising the steps of: (A) solubilizing said biological sample with a sufficient amount of a single composition to denature and solubilize said sample to form a solubilized sample, said solubilized sample comprising RNA, DNA and contaminants, wherein said composition comprises (a) at least one chaotropic agent, (b) an aromatic alcohol, and (c) a buffer present in an amount sufficient to maintain said composition at an alkaline pH; (B) extracting said solubilized sample with a water-insoluble solvent for a time sufficient to form a suspension; (C) separating said suspension into phases comprising an upper aqueous phase, a lower organic phase, and an interphase wherein said DNA is concentrated in said aqueous phase and said RNA and contaminants are concentrated in said organic phase and said interphase; and (D) precipitating said DNA from said aqueous phase and recovering said precipitated DNA.