Methods and compositions for detecting telomerase activity
US7390621B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 12, 2003 |
| Grant date | Jun 24, 2008 |
| Priority date | — |
| Expiry date | Nov 12, 2023 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/686
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for determining telomerase activity using primer extension followed with real time PCR quantification is disclosed. The method of the present invention provides a rapid, sensitive and accurate measurement for telomerase activity in a biological sample. In one embodiment, the method includes the steps of: adding the biological sample to a reaction tube containing a first reaction mixture having a first primer and nucleoside triphosphates, a second reaction mixture having a second primer and a DNA polymerase, and a wax layer that separates the first reaction mixture from the second reaction mixture; incubating the biological sample with the first reaction mixture; admixing the extension product with the second reaction mixture; amplifying and quantifying the extension product using real-time PCR and a control template. In another embodiment, the detection method includes an in situ primer extension step that allows the production of the extension product within an intact cell. In this embodiment, the extension product can be preserved under appropriate conditions for an extended time before the completion of the quantification step.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.