Patent ID: 9133503
Filing Date: 2015-09-15
Classification: C07H,C12Q

Abstract:
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.