储存
Hybridization Buffer, A⁄T1 Solution, Yeast RNA (5 mg⁄mL), RNase Digestion Buffer, RNase Inactivation⁄PPT Solution, Loading Buffer II, Mouse Kidney Total RNA, Control Template (10 µM), and Probe Elution Buffer are all stored at –20°C. Nuclease-free Water can be stored at any temperature.

描述

The Ambion® mirVana™ miRNA Detection Kit provides a fast and sensitive method for detecting small RNAs. The assay is 100–500 times more sensitive than Northern analysis, it is able to detect as little as 10 attomoles (10^-17 mol) of target RNA. Each kit includes sufficient reagents for 100 reactions.

• Sensitive—detect miRNA or siRNA in as little as 50 ng total RNA
• Specific—extremely low backgrounds
• Simple and fast—single-tube procedure eliminates the need to transfer to membrane for hybridization
• Multiple target detection—detect multiple small RNAs and mRNAs in the same sample

Rapid Procedure Based on Solution Hybridization
The mirVana™ miRNA Detection Kit is based on a simple solution hybridization principle where the RNA sample is simply mixed with radiolabeled RNA probe(s) and hybridized. Unhybridized RNA and excess probe are removed by a digestion step and the hybridized, protected RNA fragments are then recovered using a patented single-step technology for simultaneous ribonuclease inactivation and nucleic acid precipitation. RNA samples are then resuspended and analyzed on a denaturing polyacrylamide gel.

Simultaneous Detection of siRNA Expression and Target Gene Knockdown
Because multiple RNA species of the same or differing size can be detected in the same sample, the mirVana™ miRNA Detection Kit is ideal for correlating siRNA expression levels with target mRNA knockdown. An example of this application is shown in the accompanying data, which demonstrates that GAPDH mRNA levels were reduced in cells expressing a GAPDH siRNA, but not in cells expressing a control siRNA.

Accessory Products:
The mirVana™ miRNA Probe Construction Kit (SKU#AM1550) is available for the preparation and labeling of probes by in vitro transcription and the mirVana™ Probe & Marker Kit is available for 5' end labeling of oligonucleotide probes and the included Decade Markers (SKU#AM1554).

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