Amplification of midivariant DNA templates
US5407798A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Feb 5, 1993 |
| Grant date | Apr 18, 1995 |
| Priority date | — |
| Expiry date | Feb 5, 2013 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6867
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
New methods are provided for the amplification of a midivariant DNA containing an inserted target specific nucleic acid sequence(s) to enable detection of the presence of a target nucleic acid sequence in a test sample. One method employs midivariant DNA as a template for the synthesis of RNA catalyzed by QB replicase. Two midivariant DNA/probe conjugates including a nonreplicable portion of midivariant DNA and a target specific nucleic acid sequence (probe) are described. An amplification method including the steps of hybridization and ligation of the midivariant DNA/probe conjugates followed by replication of the DNA template has enabled detection of less than 200 template molecules. In a modified amplification method one of the midivariant DNA/probe conjugates further includes a RNA polymerase promoter sequence to enable transcription of the midivariant DNA template into RNA. The sequential ligation-transcription-replication method enables the detection of a single template molecule. The detectable molecule(s) are indicative of the presence of the target nucleic acid sequences in the test sample.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.