Dual phase multiplex polymerase chain reaction
US7432055B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Mar 5, 2004 |
| Grant date | Oct 7, 2008 |
| Priority date | — |
| Expiry date | Aug 25, 2024 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2565/501
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type—pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional “bridge” amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5×10−12 g (equivalent to 102-103 genomes) of a bacterial genomic DNA are disclosed.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.