Method for nucleic acid amplification by transcription using displacement, and reagents and kit therefor
US5849547A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Apr 24, 1995 |
| Grant date | Dec 15, 1998 |
| Priority date | — |
| Expiry date | Apr 24, 2015 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6846
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A target nucleic acid sequence is amplified by providing a polynucleotide such as VII including the sequence to be amplified as well as an RNA polymerase promoter sequence, and contacting said polynucleotide, in the presence of a system having the RNA polymerase activity, a RNA-dependent DNA polymerase activity, and a DNA-dependent DNA polymerase activity and capable of strand displacement, with a set of primers. The primers include a primer such as A capable of hybridizing with a segment complementary to a portion of the sequence to be amplified, wherein A includes an upstream RNA polymerase promoter sequence followed by an arbitrary sequence, a primer such as D containing said RNA polymerase promoter sequence, and primer such as C containing said arbitrary sequence. The elongation product of D is displaced by the elongation product of C, and the elongation product of D is thus obtained in the form of single-stranded VIII. Hybridization of A on VIII followed by elongation forms double stranded product X whiuch can undergo transcription, with amplification, to give single-stranded RNA VII bis. An analogous system of 3 primers B, E and F similarly produces single-stranded DNA VII fr…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.