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.
miScript miRNA PCR Array plate layout
miScript miRNA PCR Arrays are available in 96-well, 384-well, and Rotor-Disc 100 formats (see figures " Pathway-Focused or miRNome miScript miRNA PCR Array layout for plate formats A, C, D, F, M", " Pathway-Focused miScript miRNA PCR Array layout for plate formats E, G", " miRNome miScript miRNA PCR Array and miScript miRNA HC PCR Array layout for plate formats E, G", and " miScript miRNA PCR Array layout for Rotor-Disc format R"). Each array contains several control assays. The purpose of these assays is described in the table below.
Controls in miScript miRNA PCR Arrays Control | Purpose |
C. elegans miR-39 miScript Primer Assay | Alternative data normalization using exogenously spiked Syn-cel-miR-39 miScript miRNA Mimic |
6 snoRNA/snRNA miScript Primer Assays (miScript PCR Controls) | Data normalization using the ΔΔCT method of relative quantification |
miRNA reverse transcription control (miRTC) miScript Primer Assay | Assessment of reverse transcription performance |
Positive PCR control (PPC) | Assessment of PCR performance |
For more information on these controls, consult the miScript miRNA PCR Array Handbook.
miScript miRNA QC PCR Array plate layout
miScript miRNA QC PCR Arrays enable assessment of the quality of multiple cDNA samples using real-time PCR. miScript miRNA QC PCR Arrays are available in 96-well, 384-well, and Rotor-Disc 100 formats (see figures miScript miRNA QC PCR Array layout for plate formats A, C, D, F, miScript miRNA QC PCR Array layout for plate formats E and G, and " miScript miRNA QC PCR Array layout for Rotor-Disc format R"). Each miScript miRNA QC PCR Array includes replicates of C. elegans miR-39 miScript Primer Assay, 3 mature miRNA miScript Primer Assays (miR-16, miR-21, miR-191), 3 snoRNA miScript Primer Assays (SNORD61, SNORD95, SNORD96A), miRTC miScript Primer Assay, and a positive PCR control. For more information, consult the miScript miRNA PCR Array Handbook. The 96-well plate and Rotor-Disc 100 formats allow quality control of up to 8 cDNA samples. The 384-well format allows quality control of up to 32 cDNA samples.
miScript miRNA PCR Array Data Analysis Tool
Free, easy-to-use data analysis software is available online for miScript miRNA PCR Arrays. Each miScript miRNA PCR Array comes with a dedicated data analysis tool. Once raw CT values are entered, the analysis tool automatically interprets the PCR array controls, performs relative quantification using the ∆∆CT method, and presents the results in a variety of visual formats, allowing you to rapidly interpret your data.