Methods and reagents for the isolation of nucleic acids

C - Chemistry – Metallurgy – 12 – P

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Details

C12P 19/34 (2006.01) C07H 1/06 (2006.01) C07H 21/00 (2006.01) C12N 1/06 (2006.01) C12N 1/08 (2006.01) C12N 11/00 (2006.01) C12N 11/02 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2433746

The invention includes reagents and methods for the isolation of nucleic acids. The reagents described herein contain a nucleic acid precipitating agent and a solid phase carrier. The reagents can optionally be formulated to cause the lysis of a cell. These reagents can be used to isolate a target nucleic acid molecule from a cell or a solution containing a mixture of different size nucleic acid molecules. The disclosed reagents and methods provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

La présente invention concerne des réactifs et des procédés permettant l'isolement d'acides nucléiques. Les réactifs de l'invention contiennent un agent de précipitation d'acides nucléiques et un support solide. Les réactifs peuvent éventuellement être préparés de manière à provoquer la lyse d'une cellule. Lesdits réactifs peuvent être utilisés pour la mise en évidence d'une molécule cible d acide nucléique à partir d'une cellule ou d'une solution contenant un mélange de molécules d'acide nucléique de différentes tailles. Les réactifs et les procédés de l'invention fournissent un moyen simple, solide et facilement automatisable d'isolement et de purification d'acides nucléiques produisant des molécules d'acide nucléique de grande qualité aptes à l'électrophorèse capillaire, à la détermination des séquences nucléotidiques, à la transcription inverse de clonage de la transfection, à la transduction ou la micro-injection de cellules mammaliennes, aux protocoles de la thérapie génique, aux synthèses in vitro de sondes d'ARN, à la construction de bibliothèques d'ADNc et à l'amplification par la polymérase.

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