Back to results
US 5,474,916 AGranted
Promotor controlled specific amplification of nucleic acid sequences
AssigneeBoehringer Mannheim GmbH
InventorsUdo Reischl, Ruediger Rueger, Cortina Kaletta, Christoph Kessler, Joerg Kleiber
Life of this patent
Estimating the term…
Abstract
Process for the specific production of nucleic acids based on the principle of transcription in which a promoter oligonucleotide and a template-specific oligonucleotide which can hybridize with it are used as a promoter reagent and a process for nucleic acid detection which is based on this process.
Claims25
Process for the production of specific transcripts from a target nucleic acid template comprising the steps of:
- a) adding a promotor reagent P to a sample containing a target nucleic acid T, wherein at least part of nucleic acid T is in single stranded form or is converted to single stranded form, and wherein a nucleic acid complex K is formed in which the template nucleic acid is hybridized to a oligonucleotide P2 via a double-stranded region TEM 1/TEM 1' which is then hybridized to a promoter oligonucleotide P1 via a double-stranded region OBS/OBS', and
- b) forming transcripts R by promotor controlled transcription using said target nucleic acid T as a template,
- wherein the promotor reagent P contains a promotor oligonucleotide P1 and a separate template-specific oligonucleotide P2 which can hybridize with P1, and wherein the oligonucleotides P1 and P2 are not ligated together.
Process for the detection of specific nucleic acids comprising the steps of:
- a) adding a promotor reagent P to a sample containing a template nucleic acid T, wherein at least part of nucleic acid T is in single stranded form or is converted to single stranded form, and wherein a nucleic acid complex K is formed in which the template nucleic acid is hybridized to the oligonucleotide P2 via a double-stranded region TEM 1/TEM 1', which is then hybridized to a promoter oligonucleotide P1 via a double-stranded region OBS/OBS',
- b) forming transcripts R by transcription controlled by a promotor contained in P, and
- c) detecting the transcripts R, a hybrid of R and cDNA, a cDNA, a hybrid of P1 and cDNA or a mixture of these products,
- wherein the promotor reagent P contains a promotor oligonucleotide P1 and a separate template-specific oligonucleotide P2 which can hybridize with P1 and wherein the oligonucleotides P1 and P2 are not ligated together.
1 more element
Dependencies are inferred from the claim text.