Method of nucleic acid extraction
US5346999A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Mar 24, 1989 |
| Grant date | Sep 13, 1994 |
| Priority date | — |
| Expiry date | Mar 24, 2009 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10T436/25375
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
An automated apparatus is provided which implements a new method of extracting and purifying nucleic acids from cells without the use of centrifugation. In the method, a lysate is created by treating the cells with proteinase K in the presence of a lysis buffer having a high concentration of a salt. The lysate is mixed with a phenol-based solvent system, thereby creating an emulsion. The emulsion is heated to promote phase separation. Similarly, the rate of phase separation is also enhanced by increasing the surface area of the emulsion. Once the phase separation is complete, the lower organic phase is removed and the upper aqueous phase is repeatedly extracted with the phenol-based solvent a preselected number of times, and is finally extracted using chloroform. The remaining aqueous phase is then dialyzed to further purify and concentrate the nucleic acid solution. Two preferred embodiments of apparatus are presented to accomplish this extraction.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.