mirVana™ Probe & Marker Kit

Catalog number: AM1554

Ambion™  Related applications: miRNA & Non-Coding RNA Analysis

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Description

The Ambion® mirVana™ Probe & Marker Kit is designed for rapid 5' end labeling and cleanup of small RNA or DNA probes. The kit also contains reagents to prepare small, radiolabeled RNA size markers (150, 100, 90, 80, 70, 60, 50, 40, 30, 20, and 10 nt) and single-nucleotide RNA ladders. The kit includes sufficient reagents for 30 sample reactions +10 marker reactions.

Features of the Ambion® mirVana™ Probe & Marker Kit

• 5' end label and purify small RNA and DNA probes
• Includes Decade™ Markers – ideal size markers for gel analysis of small RNAs
• Simple and rapid post-labeling probe clean up procedure
• Ideal for small RNA analysis by Northern blot or with the mirVana™ miRNA Detection Kit
• Compatible with chemically synthesized oligonucleotides -—no need to gel purify after labeling

The mirVana™ Probe & Marker Kit uses an end-labeling procedure. Products from this labeling method typically do not require gel purification for subsequent use in the mirVana™ miRNA Detection Kit or RPA procedures. However, there is usually a need to remove unincorporated label. The kit includes purification columns for this purpose. Radiolabeled probes prepared with the mirVana™ Probe & Marker Kit have been successfully used for the detection of miRNA by Northern blot and by solution hybridization using the mirVana™ miRNA Detection Kit. DNA probes can also be efficiently labeled with the Probe & Marker Kit and used in many downstream applications.

How the Kit Works
Preparation of DNA and RNA probes or RNA markers starts with a phosphorylation reaction using T4 Polynucleotide Kinase and [gamma-32P]ATP followed by a rapid column purification procedure. After elution, the purified RNA is recovered in a concentrated aqueous solution. To prepare the Decade Markers, a phosphorylation reaction is performed with a 150 nt RNA transcript (included), then diluted into a cleavage reagent that generates the molecular weight marker set in a five-minute, room-temperature reaction. For exact determination of RNA size, a single ribonucleotide ladder can also be generated using any purified 5' end labeled RNA probe in a ~10 min alkaline hydrolysis reaction.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Label or Dye: 32P (Phosphorous-32)
Product Size: 1 kit
Final Product: Probes (Labeled RNA), Probes (Labeled DNA)
Labeling Method: Direct Labeling
Labeling Target: Oligos
Detection Method: Radioactive
Includes Label or Dye: No

Contents & storage

Decade™ Marker RNA should be stored at or below –70°C. T4 Polynucleotide Kinase, 10X Kinase Buffer, Control RNA Substrate, Control DNA Substrate, RNA Carrier Solution, Alkaline Hydrolysis Buffer, Loading Buffer II, and Elution Buffer are all stored at –20°C. Binding⁄Washing Buffer Concentrate should be stored at 4°C; the 10X Cleavage Reagent, Purification Cartridge + Tubes, and Elution Tubes are stored at room temperature. Nuclease-free Water can be stored at any temperature.

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