Invention Grant
- Patent Title: Allele-specific amplification of nucleic acids using blocking oligonucleotides for wild type suppression
-
Application No.: US14574181Application Date: 2014-12-17
-
Publication No.: US09909169B2Publication Date: 2018-03-06
- Inventor: Stephen G. Will
- Applicant: Roche Molecular Systems, Inc.
- Applicant Address: US CA Pleasanton
- Assignee: Roche Molecular Systems, Inc.
- Current Assignee: Roche Molecular Systems, Inc.
- Current Assignee Address: US CA Pleasanton
- Agent David J. Chang
- Main IPC: C12Q1/68
- IPC: C12Q1/68

Abstract:
A method is provided for allele-specific amplification, utilizing a blocking oligonucleotide including at least one nucleotide with a base covalently modified at the exocyclic amino group, the blocking oligonucleotide being perfectly complementary to a wild type (WT) sequence when hybridized forming a first complex having a first melting temperature (Tm), the blocking oligonucleotide being partially non-complementary, at one or more nucleotides, to a target mutant (MT) sequence when hybridized forming a second complex having a second melting temperature (Tm), wherein the first Tm is higher than the second Tm and having at least one nucleotide with a base covalently modified at the exocyclic amino group, wherein the blocking oligonucleotide becomes unhybridized from the target MT sequence during amplification but remains hybridized with the WT sequence inhibiting amplification of the WT sequence utilizing a polymerase lacking 5′-3′ nuclease activity.
Public/Granted literature
- US20160177372A1 ALLELE-SPECIFIC AMPLIFICATION OF NUCLEIC ACIDS USING BLOCKING OLIGONUCLEOTIDES FOR WILD TYPE SUPPRESSION Public/Granted day:2016-06-23
Information query