microRNA Detection by Circularization-based qRT-PCR
miR-ID and miR-Direct
microRNA Quantification by Real-Time quantitative PCR
miR-ID® : Accuracy through circularization
miR-ID® is a novel platform for detecting miRNA using a circularization-based RT-qPCR method. This technology allows SNP discrimination at any position in miRNAs and other small RNAs. miR-ID® is highly sensitive and uses affordable single-dye detection. The technology works well with all sample sources, including total RNA, cell lysates, and tissue lysates.
Circularization of linear miRNAs is followed by rolling circle amplification during the reverse transcription. The circularization step allows placement of RT primers at any location of each miRNA to optimize discriminating single nucleotide-polymorphisms (SNPs) at any location of the miRNAs.
Rolling-circle reverse transcription generates multimeric cDNAs. This method allows that primers that are entirely complementary to the miRNA sequence can be used for the real-time PCR, therefore increasing PCR specificity.
Detection of the real-time PCR signal is through SYBR chemistry.
miR-Direct® : miR-ID® - powered miRNA quantification directly from serum or plasma
SomaGenics' miR-Direct® method overcomes commonly encountered problems of inconsistent RNA recovery, PCR inhibition by sample stabilizers, and weak signals caused by low RNA abundance in biofluids. miR-Direct®- based miRNA capture is followed by miR-ID® quantification. Accurate quantification of microRNAs (miRNAs) from biofluids is crucial to realizing their potential as potential biomarkers.
miRNA is directly captured from variable input volumes (25 to 400 µl), eliminating solvent extraction or column purification. This process removes inhibitors of enzymatic reactions, including heparin.
Up to the PCR step in the miR-ID® procedure, the entire process occurs in a single tube.