Automatable Method For Nucleic Acid Isolation

Abstract

The present invention relates to a highly automatable method for isolation and / or purification of nucleic acids from a biological sample, which is particularly suitable for nucleic acids-shorter than250 bp and can be performed without a proteolytic pre-digestion step in an automated system, preferably a cartridge-based system. In a further aspect, the present invention also provides automated nucleic acid detection methods based on said isolation and / or purification method, as well as buffers and kits to be used in performing said methods.

Description

technical field [0001] The present invention relates to a highly automated method for isolating and / or purifying nucleic acids from biological samples, which is particularly suitable for nucleic acids shorter than 250 bp and can be performed in an automated system (preferably a cartridge-based system) without protein Carried out in the absence of a hydrolytic pre-digestion step. In a further aspect, the present invention also provides an automated nucleic acid detection method based on the separation and / or purification method, as well as a buffer and a kit for performing the method. Background technique [0002] Nucleic acid isolation is currently most often based on one of two different principles. The first and earlier principles employ a one-step extraction procedure whereby a buffer containing a chaotropic agent and an organic extractant (usually phenol and / or chloroform) is added to the biological sample. The mixture thus obtained thus separates into two phases: an a...

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Problems solved by technology

However, the above-mentioned procedure has a disadvantage: the above-mentioned conditions cause rapid precipitation of proteins present in the biological sample, which, without proper pretreatment, can significantly impair the adsorption of nucleic acids to the support material and also lead to The clogging of such support materials

In the presence of chaotropic agents and isopropanol, the fact that the nucleic acid-containing starting material must first be pre-digested prior to contact with the support material not only results in a protocol that is more time-consuming and laborious (which interferes with its use on automated systems efficient implementation) and importantly also increases the chances of nucleic acid degradation, especially short nucleic acids, and especially different types of RNA

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