Methods and compositions for detecting target sequences
AssigneeThird Wave Technologies, Inc.
InventorsVictor Lyamichev, Bruce P. Neri, Jeff Hall, Andrew Lukowiak
Life of this patent
Estimating the term…
Abstract
The present invention relates to compositions and methods for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. The present invention relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. For example, in some embodiments, a 5′ nuclease activity from any of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
Claims13
A method for detecting a target sequence, comprising:
- a) providing a sample containing nucleic acid suspected of containing a target sequence;
- b) generating an amplification product from said target nucleic acid, wherein said generating comprises formation of a circular nucleic acid by
- i) annealing a probe oligonucleotide that forms a first invasive cleavage structure in the presence of said target sequence,
- ii) cleaving said first invasive cleavage structure to form an annealed gapped structure,
- iii) ligating said annealed gapped structure to form a circular nucleic acid;
- c) treating said circular nucleic acid to amplification reaction conditions to produce an amplification product comprising no more than 8 contiguous bases that identically match said target sequence or its complement, and
- d) exposing said amplification product to oligonucleotides capable of forming a second invasive cleavage structure with said amplification product and an agent for detecting the presence of said second invasive cleavage structure.
Dependencies are inferred from the claim text.