Method for the simultaneous isolation of genomic DNA and high-purity RNA

US6043354A · kind A · utility

Assignee

• Invitek GmbH · DE

Inventors

• Timo Hillebrand · Spremberg - Grodk, DE
• Peter Bendzko · Berlin, DE

Key dates

• Filing date: Jul 21, 1998
• Grant date: Mar 28, 2000
• Expiry date: Jul 21, 2018

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/1006
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention concerns a method and device for the rapid, simultaneous isolation of genomic desoxyribonucleic acid (DNA) and cellular total ribonucleic acid (RNA), free of genomic DNA from various starting materials. The fields of application are molecular biology, biochemistry, gene technology (in particular gene therapy), medicine, biomedical diagnosis, veterinary medicine, food analysis and all related fields. The method proposed is characterized in that materials containing DNA and RNA are lysed in a special buffer, the lysate incubated with a mineral carrier, the carrier with the DNA bound to it separated off and washed with buffer solution, and the DNA subsequently separated from the carrier with a buffer of lower salt concentration. The lysate left after separating off the DNA bound to the carrier is mixed with phenol, chloroform and sodium acetate in defined proportions, the phases allowed to separate, and the total RNA precipitated from the aqueous phase by adding isopropanol. Lysis is carried out using buffers containing chaotropic salts with a high ionic stregth. Lysis of the material and bonding of the genomic DNA to the carrier are both carried out in the same buffer. …