Amplification of mixed sequence nucleic acid fragments
US6114149A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Aug 2, 1991 |
| Grant date | Sep 5, 2000 |
| Priority date | — |
| Expiry date | Aug 2, 2011 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6853
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method of amplifying a mixture of different-sequence DNA fragments which may be formed from RNA transcription, or derived from genomic single- or double-stranded DNA fragments. The fragments are treated with terminal deoxynucleotide transferase and a selected deoxynucleotide, to form a homopolymer tail at the 3' end of the anti-sense strands, and the sense strands are provided with a common 3'-end sequence. The fragments are mixed with a homopolymer primer which is homologous to the homopolymer tail of the anti-sense strands, and a defined-sequence primer which is homologous to the sense-strand common 3'-end sequence, with repeated cycles of fragment denaturation, annealing, and polymerization, to amplify the fragments. In one embodiment, the defined-sequence and homopolymer primers are the same, i.e., only one primer is used. The primers may contain selected restriction-site sequences, to provide directional restriction sites at the ends of the amplified fragments.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.