Qiagen, 339320, miRCURY LNA miRNA PCR Starter Kit

Two miRCURY LNA PCR Assays of your choice, spike-in control Assay (UniSp6), one candidate endogenous control assay (miR-103-3p) and all reagents for 20 reverse transcription reactions and 100 PCR amplifications; for SYBR® Green-based qPCR detection
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Features

- Easy and cost-effective introduction to the most sensitive miRNA qPCR system available
- No false positives, just true miRNA signals
- Includes RNA spike-in template and control primer set for monitoring performance
- Contains everything you need to perform small-scale experiments
- Fast and very easy protocol

Product Details

The miRCURY LNA miRNA PCR Starter Kit contains all the reagents you need to get started with the miRCURY LNA miRNA PCR System . The kit includes a spike-in control primer set (UniSp6), one candidate endogenous control primer set (miR-103a-3p) and two validated primer sets of your choice, enhanced with LNA technology . The reagents are sufficient for performing 20 reverse transcription reactions and 200 PCR amplifications, enabling quantification of up to 20 samples with a minimum of 4 assays per sample. Want to try this solution for the first time? Request a quote for a trial kit .

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Performance

Unmatched sensitivity

Universal reverse transcription combined with LNA-enhanced and T m -normalized primers enables accurate and reliable quantification of individual miRNAs from as little as 1 pg total RNA (see figure Accurate quantitation from down to 1 pg total RNA starting material ). With exceptional sensitivity and extremely low background, you can accurately quantify very low levels of miRNA without pre-amplification. This makes the miRCURY LNA miRNA PCR System suitable for all types of samples, especially those with low RNA content, such as serum, plasma and other biofluids.

Principle

High-performance reagents for quantitative, real-time RT-PCR

miRCURY LNA RT Kit This kit includes all the necessary reagents to perform fast and convenient miRNA polyadenylation and reverse transcription in a single reaction step. Using a single, universal RT reaction as the template for subsequent PCR amplifications greatly simplifies the procedure compared to systems that require miRNA-specific, first-strand synthesis (see figure The benefit of a universal reverse transcription step ). The UniSp6 RNA spike-in can be used with the control assay – UniSp6 miRCURY LNA miRNA PCR Assay (cat. no. YP00203954) – as a reverse transcription positive control and/or as an inter-plate calibrator in our miRNA PCR panels. miRCURY LNA SYBR Green PCR Kit This kit contains the high-performance miRCURY SYBR Green Master Mix for quantitative real-time PCR amplification. The detection is based on SYBR Green, which allows quality control of the resulting PCR amplicon by melting curve analysis. LNA-enhanced primer sets require specific PCR reaction conditions for optimal performance. The kit also includes a control primer set designed for amplification of the synthetic RNA spike-in provided in the miRCURY LNA RT Kit. Refer to the miRCURY LNA RT Kit Handbook for details. The miRCURY LNA miRNA PCR Assays and Panels have been validated using the miRCURY SYBR Green Master Mix; use of other reagents for qPCR will affect the results.

Procedure

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.

Applications

- Setup of miRNA quantification experiments
- miRNA control experiments

The miRCURY LNA miRNA PCR Starter Kit is used as part of the miRCURY PCR System for: