miRNA expression profiling with miScript miRNA PCR Arrays is straightforward and robust. First, prepare cDNA buffered with miScript HiSpec Buffer using the miScript II RT Kit. Second, add a premix of cDNA, miScript Universal Primer, QuantiTect SYBR Green PCR Master Mix, and RNase-free water to a miScript miRNA PCR Array. Third, run the reaction in a real-time PCR cycler. Finally, analyze the data using the miScript miRNA PCR Array Data Analysis Tool (see figure miScript miRNA PCR Array workflow
Designing your Custom miScript miRNA PCR Array
1. Select a plate layout to match the miRNA number and the number of samples you would like to analyze (see figure Plate layouts
2. Determine the number of plates required for your experiment.
3. Use the Excel file to enter your miRNA list. Supply the mature miRNA ID and miRBase accession number for each miRNA. (Visit GeneGlobe or miRBase to find the miRBase accession numbers for your miRNAs of interest).
4. Include the type of PCR cycler to be used for the profiling.
5. Contact Technical Service to submit your miRNA list and obtain a quote.