Methods and compositions for amplification and capture of nucleic acid sequences
US7501254B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Jul 19, 2007 |
| Grant date | Mar 10, 2009 |
| Priority date | — |
| Expiry date | Aug 8, 2027 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6846
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods for amplification and capture of nucleic acid sequences include annealing a forward primer to a DNA or RNA template in a first reaction vessel including fewer than four different dNTPs; forming an extended primer that terminates when an omitted dNTP is required for further extension; releasing the extended primer; exponentially amplifying the extended primer in a second reaction vessel that includes a reverse primer, four different dNTPs and a capture probe that includes n oligonucleotides having fewer than n locking nucleic acids; and concurrently capturing one of the extended primers with the capture probe while amplifying the extended primer. The steps of annealing, extending and releasing can occur at a first reaction temperature that is substantially isothermal and in the absence of a helicase. The steps of exponentially amplifying and capturing can occur at a second reaction temperature that is substantially isothermal.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.