描述
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! 规格
Detection Method: | SYBR |
PCR Method: | 2-Step RT-PCR, 2-Step qRT-PCR |
Polymerase: | SuperTaq™ Polymerase (Not Included) |
Product Line: | mirVana™, Ambion™ |
Product Size: | 200 reactions |
Sample Type (General): | RNA |
Sample Type (Specific): | Total RNA, miRNA |
For Use With (Equipment): | 7500 System, 7900HT System, StepOne™, StepOnePlus™, ViiA™ 7 System |
Shipping Condition: | Dry Ice |
内容及储存
440 µL mirVana™ 5X RT Buffer –20°C
1.1mL 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 1.75mL Nuclease-free Water - any temp