Method of synthesizing cDNA
US8067205B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Mar 29, 2004 |
| Grant date | Nov 29, 2011 |
| Priority date | — |
| Expiry date | Sep 30, 2024 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1096
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3′ and 5′ ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.