mirVana™ qRT-PCR miRNA Detection Kit

Catalog number: AM1558

Ambion™  Related applications: Real Time PCR (qPCR) | miRNA & Non-Coding RNA Analysis

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Description

The mirVana™ qRT-PCR Detection Kit has been optimized to amplify specific small RNA using qRT-PCR primer sets (not included), enabling accurate quantitation of miRNA expression levels. The novel detection system can be used to characterize miRNA expression via traditional endpoint RT-PCR or real-time RT-PCR using SYBR® Green I.

• Quantitative results in less than 3 hours
• Detection from as little as 25 pg of total RNA
• Suitable for either real-time or end-point RT-PCR
• Ideal for validating expression profiling data and high-throughput applications.

MicroRNAs (miRNAs) are evolutionarily conserved, small, non-coding RNA molecules that are estimated to regulate at least 30% of all protein encoding genes. Currently more than 2,900 miRNAs have been annotated from a wide range of organisms including plants, worms, flies, mammals, and viruses. To date, miRNAs have been implicated in development, differentiation, apoptosis, fat metabolism, viral infection, and cancer.

Due to their small size (~22 nt), miRNAs cannot be analyzed by traditional PCR approaches. Designed for use in conjunction with qRT-PCR Primer Sets (not included), the mirVana™ qRT-PCR Detection Kit has been optimized to amplify specific small RNAs, enabling accurate quantitation of miRNA expression levels (Figure 1). This novel detection system can be used to characterize miRNA expression via traditional endpoint RT-PCR or real-time RT-PCR using SYBR® Green I.

Since miRNAs represent only a minute fraction of total RNA, detection of these molecules can be difficult. However, the mirVana™ qRT-PCR miRNA Detection Kit is extremely sensitive, detecting miRNA from as little as 25 pg of total RNA. The kit also specifically detects mature miRNAs and not miRNA precursors (Figure 2). Furthermore, the mirVana™ qRT-PCR miRNA Detection Kit exhibits linearity over at least 5 logs (Figure 3) and can be used with total RNA, miRNA-enriched RNA fractions, or even degraded RNA samples.

Each kit contains sufficient reagents for 200 reactions, as well as a control primer set to amplify the ubiquitously expressed miR-24 and human heart total RNA. Note: Thermostable DNA polymerase is not supplied with the kit. For analysis of individual miRNAs, the mirVana™ qRT-PCR miRNA Detection Kit is unparalleled in sensitivity and speed!
For Research Use Only. Not for use in diagnostic procedures.

Specifications

PCR Method: 2-Step qRT-PCR, 2-Step RT-PCR
Polymerase: SuperTaq™ Polymerase (Not Included)
Product Size: 200 reactions
Detection Method: SYBR
Sample Type (General): RNA
Sample Type (Specific): Total RNA, miRNA
For Use With (Equipment): 7900HT System, 7500 System, ViiA™ 7 System, StepOnePlus™, StepOne™
Shipping Condition: Dry Ice

Contents & storage

440 µL mirVana™ 5X RT Buffer –20°C
1.1 ml mirVana™ 5X PCR Buffer –20°C
88 µL ArrayScript™ RT Enzyme Mix –20°C
22 µL Human Heart Total RNA (25 ng⁄µL) –20°C
10 µL 10X has-miR-24 RT Primer –20°C
11 µL has-miR-24 PCR Primers –20°C
2 x 1.75 mL Nuclease-free Water - any temp

Documents

Manuals & protocols