miRCURY LNA RT Kit

• Optimized for both detection methods of miRCURY LNA miRNA PCR Assays and Panels
• Fast, easy and reliable first-strand synthesis
• Includes RNA spike-in template for monitoring performance

The miRCURY LNA RT Kit enables fast and convenient miRNA polyadenylation and reverse transcription in a single reaction step. Using one universal RT reaction as the template for subsequent PCR amplifications greatly simplifies the procedure compared with systems that require miRNA-specific first-strand synthesis, adding convenience and saving you time. The kit has been designed for use with the two miRCURY LNA miRNA PCR Systems, using either SYBR Green or an LNA-enhanced hydrolysis probe as the detection method. The kit contains two separate reaction buffer tubes for the respective PCR detection method.

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Simplified reverse transcription saves time and resources

Using a universal reverse transcription reaction per RNA sample saves not only reagents and precious samples, but also time. The miRCURY LNA RT Kit is used in first-strand cDNA synthesis for both miRCURY LNA miRNA PCR Systems, employing SYBR Green for detection (see  miRCURY LNA miRNA PCR System at a glance) or an LNA-enhanced, miRNA-specific hydrolysis probe (see miRCURY LNA miRNA Probe PCR System at a glance). The kit includes a reaction buffer that is specific to each system. The miRNA assays and panels are system-specific.

Unmatched sensitivity

Universal reverse transcription combined with LNA-enhanced and T_m-normalized PCR components 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 and the miRCURY LNA miRNA Probe PCR System suitable for all types of samples, especially those with low RNA content, such as serum, plasma and other biofluids.

Fast and Easy

The miRCURY LNA RT Kit is the first step for the miRCURY LNA miRNA PCR Systems, using either SYBR Green or a hydrolysis probe for detection. The easy-to-follow, 2-hour protocol saves time and labor. By using a universal reverse transcription as well as optimized RT-PCR protocols, the procedure is greatly simplified compared to other solutions. Furthermore, the cDNA generated in the RT step can be used across your chosen system, allowing you to seamlessly transition from panels to assays depending on your research needs, and saving sample material and time. The number of pipetting steps is reduced to a minimum, which also minimizes technical variation. This simplified workflow coupled to ISO-compliant assay manufacturing makes it possible to achieve extremely high day-to-day and site-to-site reproducibility.

One miRCURY LNA RT Kit for two detection systems

The miRCURY LNA RT Kit is used for both miRCURY LNA miRNA PCR Systems, either SYBR Green or hydrolysis probe detection methods. This kit includes two system-specific reaction buffers and 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). Furthermore, the cDNA generated in the RT step can be used across your chosen system, allowing you to seamlessly transition from panels to assays depending on your research needs and saving sample material and time. The UniSp6 RNA spike-in included in the kit can be used with the control assay – UniSp6 miRCURY LNA miRNA PCR Assay – as a reverse transcription positive control and/or as an inter-plate calibrator in our miRNA PCR panels.

miRCURY LNA miRNA PCR System (SYBR Green)

The architecture of this system consists of SYBR Green detection with two LNA-enhanced and miRNA-specific primers for unmatched performance (see miRCURY LNA miRNA PCR System at a glance).

• miRCURY LNA RT Kit
• RNA Spike-In Kit
• miRCURY LNA SYBR Green PCR Kit
• miRCURY LNA miRNA PCR Assays (predefined, Custom)
• miRCURY LNA miRNA PCR Panels (miRNome, Focus, Custom)

miRCURY LNA miRNA Probe PCR System

A unique LNA-enhanced, miRNA-specific hydrolysis probe is combined with an LNA-enhanced, miRNA-specific forward primer for highly specific and sensitive true signals (see miRCURY LNA miRNA Probe PCR System at a glance).

• miRCURY LNA RT Kit
• RNA Spike-In Kit
• miRCURY LNA Probe PCR Kit
• miRCURY LNA miRNA Probe Assays (predefined, Custom)
• miRCURY LNA miRNA Probe Panels (miRNome, Focus, Custom)

Reverse transcription reactions are easy to perform as single-tube reactions. Total RNA, including miRNA, is used as starting material and the procedure involves incubating the reaction at 40–42°C for 1 hour, followed by an inactivation step though a brief incubation at 95°C.

The miRCURY LNA RT Kit is used as part of the miRCURY LNA miRNA PCR and Probe PCR Systems for:

• miRNA expression detection, profiling and quantification
• miRNA biomarker development – screening and identification
• miRNA regulation and pathway analysis
• miRNA detection in complex sample types and samples with low miRNA expression levels
• snoRNA detection
• Validation of miRNA sequencing