Methods for using double-mutant RNA polymerases with reduced discrimination between non-canonical and canonical nucleoside triphosphates
US7309570B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Oct 20, 2003 |
| Grant date | Dec 18, 2007 |
| Priority date | — |
| Expiry date | Jan 4, 2025 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6869
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method and kit for synthesizing a nucleic acid molecule comprising at least one non-canonical nucleoside triphosphate using a double-mutant polymerase having a reduced discrimination between canonical and non-canonical substrates is disclosed. The method comprises incubating a template nucleic acid in a reaction mixture comprising the mutant nucleic acid polymerase and the appropriate canonical and non-canonical nucleoside triphosphates which are desired substrates for the mutant nucleic acid polymerase. The present invention is also a method of determining the sequence of a nucleic acid molecule using the mutant polymerase to create a nucleic acid molecule comprising at least one non-canonical nucleoside triphosphate.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.