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Background/Introduction: The miRNAs RT-PCR Quantitation Kit is a sensitive and specific method using real-time PCR for the detection and quantification of miRNAs (miRNA) from total RNA samples. miRNAs are small, single-stranded, ~19-23 nt RNA molecules encoded in the genomes of plants, animals, and viruses. Mature miRNAs enter the RNA-induced silencing complex (RISC) and guide the RISC to induce translational repression or endonucleolytic cleavage of specific target mRNAs. Unlike commonly used methods for detection of miRNAs, the miRNAs qPCR Quantitation Kit is more rapid and sensitive. This kit contains miRNA hsa-mir-126-3p and cel-mir-39-3p high specific RT and PCR primer set, with SYBR Green dye included. The stem-loop like miRNAs RT primer and the miRNAs high specific primer set ensure the RT and PCR reaction would not be interfered by the miRNAs precursors. The reaction template could be total RNA or cell lysates. The system enables highly sensitive detection from as few as 10 copies of a target miRNAs, with a broad dynamic range that supports accurate quantification of high-copy mRNA from up to 1 ug of total RNA. High specificity ensurenced by stemloop RT primer and high specific miRNAs primer set; even highly homogized miRNAs can be accurately dicriminated. Higher dynamic range and sensitivity of miRNAs quantification than conventional method (such as Northern blot & micro-array) with broad dynamic range of at least seven orders of magnitude, and in this ready to use formulation can quantify as few as 7 copies of a miRNA target in as little as 0.1 pg of total RNA. Less sample requirement Total RNA, cell lysate or purified samll RNA can work as the qPCR Quantitation Kit's template, even the genomic DNA contamination would not interfere in miRNAs quantification. miRNA synthetic standard can work as a standard curve when you need to know the absolute number of a miRNA in one cell or work as positive control when you need to know the relative expression ratio between miRNA and cel-mir-39-3p or 5S rRNA.
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