Inhibition of mismatch hybridization by a universal competitor dna
AssigneeGeneOhm Sciences, Inc.
InventorChunnian Shi
Life of this patent
Estimating the term…
Abstract
Provided herein are compositions and methods for enhancing the relative efficiency of hybridization between target nucleic acids and capture probes compared to target variants and capture probes.
Claims47
A method for increasing hybridization efficiency between a capture probe and a target nucleic acid, comprising:
- providing a probe complex comprising a capture probe and a linker sequence, wherein said capture probe comprises a complementary region comprising a nucleic acid that is substantially complementary to a target nucleic acid sequence, and wherein said linker sequence is operably linked to said capture probe;
- providing a target nucleic acid, wherein said target nucleic acid or a fragment thereof is substantially complementary to said complementary region of said capture probe;
- providing a competitor nucleic acid, wherein said competitor nucleic acid comprises a complementary region that is fully complementary to said linker sequence and a universal region comprising at least about two universal bases that can hybridize to A, T, C and G, wherein said universal region is adjacent to the 5′ end of said complementary region of said competitor nucleic acid;
- and
- contacting said probe complex with said target nucleic acid and said competitor nucleic acid.
The method of claim 6, wherein said at least two universal bases are deoxyinosine.
A method of increasing the sensitivity and specificity of a nucleic acid hybridization assay, comprising:
- providing a capture probe that is substantially complementary to a target nucleic acid sequence in a sample;
- providing a competitor nucleic acid that reduces the hybridization efficiency of non-target nucleic acids to said capture probe;
- and
- contacting said capture probe with said sample and said competitor nucleic acid.
A competitor nucleic acid to increase hybridization efficiency between a probe complex and a target nucleic acid, wherein said probe complex comprises a complementary region comprising a nucleic acid that is complementary to a target nucleic acid sequence and a linker sequence, wherein said capture probe and said linker sequence are operably linked, comprising
- a complementary region that is substantially complementary to said linker sequence, and
- a universal region comprising at least about two universal bases, wherein said universal region is adjacent to the 5′ end of said complementary region of said competitor nucleic acid.
The competitor nucleic acid of claim 45, wherein said universal base is selected from the group consisting of deoxyinosine, 3-ntiropyrorole, 4-nitroindole, 6-nitroindole, and 5-nitroindole.
The competitor nucleic acid of claim 45, wherein said universal base is deoxyinosine.
Dependencies are inferred from the claim text.