Kits for detection of nucleic acids
First Claim
1. A kit for detecting an miRNA target comprising a first region, a second region, and a third region, wherein said first region is downstream of and contiguous to said second region, and wherein said second region is downstream of and contiguous to said third region, comprising:
- i) a first oligonucleotide having a 3′
end, wherein said first oligonucleotide comprises a 3′
portion that is complementary to said first region of said miRNA target, and a 5′
portion that is not complementary to said miRNA target, wherein said 3′
portion of said first oligonucleotide comprises the 3′
end of said first oligonucleotide;
ii) a second oligonucleotide having a 3′
end, wherein said second oligonucleotide comprises a 3′
portion that is substantially homologous to said third region of said miRNA target and a 5′
portion that is not homologous to said third region of said miRNA target, wherein said 3′
portion of said second oligonucleotide comprises the 3′
end of said second oligonucleotide;
iii) a reverse transcriptase;
iv) a DNA polymerase;
v) a 5′
nuclease;
vi) a probe oligonucleotide, wherein at least a portion of said probe oligonucleotide is complementary to at least a portion of said second region of said miRNA target; and
vii) a first stacker oligonucleotide comprising a 5′
terminal portion, wherein at least the 5′
terminal portion of said stacker oligonucleotide is complementary to a region in said 5′
portion of said first oligonucleotide adjacent to said 3′
portion of said first oligonucleotide.
7 Assignments
0 Petitions
Accused Products
Abstract
The present invention relates to compositions and methods for the detection and characterization of small nucleic acid molecules (e.g., RNA (e.g., small RNAs such as micro RNAs (miRNAs) and small interfering RNAs (siRNAs)) and other short nucleic acid molecules). More particularly, the present invention relates to methods for the detection and quantification of RNA expression. The present invention further provides for the detection of miRNA and siRNA variants.
83 Citations
14 Claims
-
1. A kit for detecting an miRNA target comprising a first region, a second region, and a third region, wherein said first region is downstream of and contiguous to said second region, and wherein said second region is downstream of and contiguous to said third region, comprising:
-
i) a first oligonucleotide having a 3′
end, wherein said first oligonucleotide comprises a 3′
portion that is complementary to said first region of said miRNA target, and a 5′
portion that is not complementary to said miRNA target, wherein said 3′
portion of said first oligonucleotide comprises the 3′
end of said first oligonucleotide;ii) a second oligonucleotide having a 3′
end, wherein said second oligonucleotide comprises a 3′
portion that is substantially homologous to said third region of said miRNA target and a 5′
portion that is not homologous to said third region of said miRNA target, wherein said 3′
portion of said second oligonucleotide comprises the 3′
end of said second oligonucleotide;iii) a reverse transcriptase; iv) a DNA polymerase; v) a 5′
nuclease;vi) a probe oligonucleotide, wherein at least a portion of said probe oligonucleotide is complementary to at least a portion of said second region of said miRNA target; and vii) a first stacker oligonucleotide comprising a 5′
terminal portion, wherein at least the 5′
terminal portion of said stacker oligonucleotide is complementary to a region in said 5′
portion of said first oligonucleotide adjacent to said 3′
portion of said first oligonucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
-
Specification