Methods and compositions for detecting differences between nucleic acids
AssigneePanomics, Inc.
InventorQinghong Yang
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Abstract
The present invention provides methods for detecting the presence or absence of a difference between two related nucleic acid sequences. The methods achieve sensitivities great enough to detect the presence of any difference between the nucleic acids, even single nucleotide polymorphisms. In the methods, the nucleic acids are contacted under conditions in which they are capable of forming a stable four-way complex if the nucleic acids differ in sequence. The presence of the four way complex is determined with a detection molecule and a labeled four-stranded tracer molecule.
Claims26
A method for detecting a difference in the sequence of two nucleic acid molecules comprising:
- a. Contacting said two nucleic acids under conditions that allow the formation of a four-way complex and branch migration;
- b. Contacting said four-way complex with a tracer molecule and a detection molecule under conditions in which the detection molecule is capable of binding the tracer molecule or the four-way complex;
- and
- c. Determining binding of tracer molecule to the detection molecule before and after exposure to said four-way complex, wherein reduced binding after contact indicates the presence of a difference between the two nucleic acids.
A method for detecting a difference in the sequence of two nucleic acid molecules comprising:
- a. Contacting said two nucleic acids under conditions that allow the formation of a four-way complex and branch migration;
- b. Contacting said four-way complex with a tracer molecule and a detection molecule under conditions in which the detection molecule is capable of binding the tracer molecule or the four-way complex;
- and
- c. Comparing binding of said tracer molecule said detection molecule in b. with binding of said tracer molecule to said detection molecule in said test solution without said four-way complex, wherein reduced binding in b. indicates a difference between the two nucleic acids.
A tracer molecule comprising a detectable label and a nucleic acid complex, wherein said nucleic acid complex comprises a stable four-way junction.
Dependencies are inferred from the claim text.