miRCURY PCR Controls
Validated primer sets for normalization of miRNA expression levels in real-time PCR analyses
To obtain accurate and reproducible results in real-time PCR-based miRNA quantification, it is necessary to normalize the amount of target miRNA using a suitable endogenous reference RNA, a process known as relative quantification. miRCURY PCR Controls are primers that enable normalization of real-time PCR results in miRNA quantification studies using the miRCURY LNA miRNA PCR system.
miRCURY PCR Controls are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Reference genes for normalization
miRCURY LNA miRNA PCR reference genes are constitutively and relatively stable expressed across different human tissues (see figure Expression levels of reference genes in different human tissues). The reference genes cover a large range of Cq values, offering the possibility to accurately and reliably normalize across a range of miRNA expression levels.
A unique system for miRNA profiling
miRCURY LNA miRNA PCR Assays offer the best combination of performance and ease-of-use available on the miRNA real-time PCR market by combining universal RT with LNA PCR amplification. Universal RT makes it possible to use one first-strand cDNA synthesis reaction as the template for multiple miRNA real-time PCR assays. This saves precious samples, reduces technical variation and saves time in the laboratory. Plus, both the forward and reverse PCR amplification primers are miRNA specific and optimized with LNA. This provides 1) exceptional sensitivity and extremely low background, enabling accurate quantitation of very low miRNA levels and 2) highly specific assays that allow discrimination between closely related miRNA sequences.
Reference gene assays for normalization
Nine reference gene primer sets are available, enabling high-quality data normalization and generation of reliable data from many different sample types (see table below). These control primer sets target endogenous, small non-coding RNAs that are constitutively and relatively stably expressed across different human tissues (see figure Expression levels of reference genes in different human tissues). The control reference genes cover a large range of Cq values, offering the possibility to accurately and reliably normalize across a range of miRNA expression levels.
The control primer sets have been validated as reference genes for the miRCURY LNA miRNA PCR System and work optimally with the miRCURY LNA RT Kit and the miRCURY LNA SYBR Green PCR Kits. It is very important to always confirm the consistent and unvarying expression of a gene in all tissues and under all treatments of an experiment before choosing it as the reference. We recommend that you test a number of reference genes and use software such as GeNorm and NormFinder (part of the GenEx software) to choose and validate reference genes.
List of available reference gene assays
The miRCURY LNA miRNA PCR Assay system is an miRNA-specific, LNA-based system designed for sensitive and accurate detection of miRNA by quantitative real-time PCR using SYBR Green. The first step of the procedure is universal reverse transcription, followed by real-time PCR amplification with LNA-enhanced primers.
miRCURY LNA miRNA PCR Assays are used as part of the miRCURY LNA miRNA PCR System for:
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