Method for optimized isolation of RNA from fixed tissue
US9051603B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 8, 2009 |
| Grant date | Jun 9, 2015 |
| Priority date | — |
| Expiry date | Sep 1, 2032 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1003
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
In invention relates to a method for the isolation of RNA from tissue pretreated with formaldehyde comprising homogenizing the sample in the presence of a guanidinium salt in aqueous solution, and incubating the sample in the presence of 0.1 M to 5 M ammonium salt at a temperature between 50° C. and 100° C. The heat treatment in the presence of an ammonium salts demodifies RNA by reverting methylol groups which are formed in the presence of formaldehyde between amino groups in nucleobases of RNA and in basic amino acids, and by cleavage of methylene bridges between amino groups in nucleobases of RNA and basic amino acids, to provide high RNA recoveries and consistently high quality of RNA for further reaction, e.g. for reverse transcriptase-polymerase chain reaction or microarray analysis.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.