miScript miRNA PCR Arrays

For SYBR® Green-based, real-time PCR profiling of miRNAs using the miScript PCR System

Features

  • Sensitive and specific mature miRNA expression profiling
  • PCR arrays for pathway, disease, or miRNome profiling
  • Free online data analysis tools

Products

miScript miRNA PCR Arrays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
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miRNome V16

Cat. No. / ID: 331222

miRNome panels of miRNA assays in 96-well, 384-well, or Rotor-Disc formats
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Pathway-Focused miScript miRNA PCR Array

Cat. No. / ID: 331221

Pathway- or disease-focused panels of miRNA assays in 96-well, 384-well, or Rotor-Disc formats
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miScript miRNA HC PCR Array

Cat. No. / ID: 331223

High-content array of assays for a pathway, disease, or gene family miRNAs; available in 384-well format
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miScript miRNA QC PCR Array

Cat. No. / ID: 331292

Array of quality control assays available in 96-well, 384-well, or Rotor-Disc format

Product Details

miScript miRNA PCR Arrays are mature miRNA-specific forward primers (miScript Primer Assays) that have been arrayed in biologically relevant pathway-focused, disease-focused, or whole miRNome panels. miScript miRNA PCR Arrays are part of the miScript PCR System for miRNA detection and quantification. cDNA is prepared using the miScript II RT Kit with miScript HiSpec Buffer and is used as a template in real-time PCR with a miScript miRNA PCR Array and the miScript SYBR Green PCR Kit.

Important note: This product line will be discontinued on May 1, 2021 and will be replaced by the miRCURY LNA miRNA PCR system. We strongly recommend using the following miRCURY LNA PCR products for better miRNA quantification and profiling: miRNome PCR Panels, miRNA Focus PCR Panels and miRNA QC PCR Panel.

For more information about this transition, please visit our dedicated knowledge hub or contact us.

Performance

Discover biomarkers in serum or plasma samples

The range of pathway-focused miScript miRNA PCR Arrays is continually expanding to enable new discoveries about the roles of miRNAs in biological processes. The content of miScript miRNA PCR Arrays is selected using our proprietary methodology, which ensures that each array is up-to-date and biologically relevant. One of the most exciting areas of current miRNA research involves the assessment of miRNAs present in serum or plasma samples. The discovery of relatively stable, extracellular miRNAs in serum and plasma has generated huge interest in the use of changes in these miRNA levels as potential noninvasive biomarkers for a variety of diseases. As a result, the Serum & Plasma miScript miRNA PCR Array has been developed. This array enables rapid profiling of the 84 most relevant miRNAs associated with serum and plasma (see figure " Analysis of expression of circulating miRNAs in serum or plasma").

Profile entire miRNomes with a single cDNA sample

In addition to pathway-focused arrays, up-to-date miRNome miScript miRNA PCR Arrays are available for a variety of species including human, mouse, rat, dog, and rhesus macaque. With less than 1 µg RNA, the entire human miRNome can be rapidly profiled (see figure " Whole miRNome profiling"). Each assay in a miScript miRNA PCR Array has been bench verified to ensure sensitive and specific detection of mature miRNA by real-time PCR.

See figures

Principle

The miScript PCR System enables sensitive, specific miRNA quantification and profiling using SYBR Green real-time PCR. The miScript PCR System covers all the steps involved in conversion of RNA to cDNA and subsequent real-time PCR detection of miRNAs.

Mature miRNA profiling with the miScript PCR System uses the following components:

  • miScript II RT Kit — this kit enables simple, single-step cDNA synthesis. A single cDNA synthesis reaction can be used for detection of hundreds of miRNAs. The dual buffer system meets the distinctive needs of miRNA quantification using real-time PCR.
  • miScript SYBR Green PCR Kit — this kit includes QuantiTect SYBR Green PCR Master Mix and the miScript Universal Primer, a reverse primer that allows detection of miRNAs in combination with a miScript Primer Assay or miScript miRNA PCR Array.
  • miScript miRNA PCR Array — Cutting-edge miScript miRNA PCR Arrays consist of mature miRNA-specific forward primers (miScript Primer Assays) arrayed in biologically relevant pathway- or disease-focused panels or miRNome panels. 

Procedure

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.

See figures

Applications

miScript miRNA PCR Arrays are used as part of the miScript PCR System for:

  • Pathway- and disease-focused miRNA profiling
  • miRNome profiling
  • Biomarker discovery

Supporting data and figures

Resources

Brochures & Guides (4)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
Kit Handbooks (2)

miScript II RT Kit

miScript SYBR Green PCR Kit

miScript miRNA PCR Arrays

miScript miRNA QC PCR Array

For SYBR Green-based, real-time PCR profiling of microRNAs using pathway-focused arrays, HC arrays, and miRNome arrays

Instrument Technical Documents (2)
For gene expression and genomic analysis
For pathway-focused miRNA or miRNome analysis
Download Files (1)
Data analysis file for miScript miRNA PCR Array All miRNA QC (96-well plate, Rotor-Disc 100)
Catalog number- 331221
Pathway number- MIXX-989Z

FAQ

Can miScript miRNA qPCR Assays detect single nucleotide difference between miRNAs?
Yes. With our patent pending proprietary technology, miScript miRNA qPCR Arrays and Assays can distinguish miRNAs that differ by only one single nucleotide.
FAQ ID -2723
Will pipetting error affect the miRNA qPCR Assay results?
The passive reference dye, ROX in the miScript PCR master mix is used by real-time PCR systems to normalize variation from well to well. Therefore, these systems tolerate volume variations caused by pipetting error and evaporation.
FAQ ID -2728
Do I need the sequence specific miRNA primers in the RT step of the miRNA qPCR Assays?
No. Because the miScript miRNA qPCR assay technology uses universal reverse transcription in the RT step, sequence specific miRNA primers are not needed in this step. Therefore, cDNA obtained from universal RT can be used for any miScript miRNA expression analysis qPCR assay or PCR array.
FAQ ID -2725
How can I prevent the evaporation of reaction volume from the wells in the miScript miRNA PCR array?
Be sure to carefully and completely seal the PCR plate with the optical thin-wall 8-cap strips or the optical adhesive film before placing it into your thermal cycler. Also, be sure to use a compression pad when using an optical film seal as directed by the manufacturer of your real-time PCR instrument.
FAQ ID -2729
Why did the real-time PCR yield Ct values < 12?
You probably used too much template. Try less input RNA for reverse transcription especially if the higher end of the recommended range had been used previously. Or use a smaller volume of template per PCR reaction or use diluted template, but do not use less than 1 µl of template per each 25-µl miRNA PCR array reaction. Remember to use the same volume of template in each reaction.
FAQ ID -2727
Can miScript miRNA qPCR Assays be used for pre-miRNA detection?
No. miScript miRNA PCR Arrays and miScript Primer Assays are designed to measure exclusively the expression of mature miRNA, not pre-miRNA. To measure miRNA precursors use miScript Precursor Assays available through the QIAGEN GeneGlobe assay search portal.
FAQ ID -2724
Can I use total RNA for the miRNA PCR Arrays or Assays?
Yes, you can. In fact, total RNA is the recommended starting material for the miScript PCR System. We recommend using the miRNeasy Mini Kit (217004) to isolate total RNA for use with the miScript PCR System.
FAQ ID -2726