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Features
- Optimized for use with miRCURY LNA miRNA PCR Assays and Panels
- Sensitive and specific detection and quantification of miRNA
- Includes optional ROX dye for optional normalization of fluorescence
Product Details
The miRCURY LNA SYBR Green PCR Kit contains a high-performance PCR master mix specifically designed for use with the miRCURY LNA miRNA PCR System. The miRCURY SYBR Green PCR Kit also features a built-in control for visual identification of correct template addition, as well as Q-Bond, an additive in the PCR buffer that enables short cycling steps without loss of PCR sensitivity and efficiency. The kit has been optimized for use with any real-time cycler. ROX Reference Dye is provided in a separate tube and can be added if your cycler requires ROX as a passive reference dye.
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Performance
Unmatched sensitivity
Universal reverse transcription combined with LNA-enhanced and Tm-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.
See figures
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.
See figures
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 (see figure miRCURY LNA miRNA PCR System at a glance).
See figures
Applications
The miRCURY LNA SYBR Green PCR Kits are used as part of the miRCURY PCR System for:
- Mature miRNA quantification and profiling
- Mature miRNA and precursor miRNA detection in parallel
- snoRNA RNA detection
Supporting data and figures
Accurate quantitation from down to 1 pg total RNA starting material.
Data from serial dilutions of human AM6000 total RNA are shown. All miRNA assays exhibit linear readout with correlation coefficients R2 > 0.99.
Resources
Safety Data Sheets (1)
Quick-Start Protocols (2)
Scientific Posters (1)
Additional Resources (1)
Brochures & Guides (2)
Kit Handbooks (2)
Certificates of Analysis (1)