Overview
The most advanced RNA tracking, visualisation and pull down technology. Technology for studying the diverse cellular roles of RNA has lagged behind the tools for studying DNA and proteins, but innovative researchers are working to change that! One such researcher is Dr. Peter Unrau of Simon Fraser University. He and his team have created RNA Mango, a novel technology with a number of useful applications.
Products Available:
| Product Name | Cat. No. | Size |
|---|---|---|
| TO1-3PEG-Biotin Fluorophore | ABM-G7955 | 250 µM (100 µl) |
| TO1-3PEG-Desthiobiotin Fluorophore | ABM-G7956 | 250 µM (100 µl) |
| TO3-3PEG-Biotin Fluorophore | ABM-G7959 | 250 µM (100 µl) |
| YO3-3PEG-Biotin Fluorophore | ABM-G7957 | 250 µM (100 µl) |
* Click here for data on binding affinity to Mango and Peach aptamers
Key Features
RNA Mango technology is based on the specific binding of the RNA Mango Aptamer and a Thizole Orange (TO) bi-functional dye. The main features of this technology is the tight binding between the dye and aptamer (KD ≈ 3nM) , and the strong ~1000X enhancement of the dye’s fluorescence when bound to the Mango aptamer (Fluorescent enhancement FE=1,100).
The Thizole Orange (TO) dye has a number of other desirable properties including:
- Small size
- Lack of toxicity
- Plasma and nuclear membrane permeability
- Short intracellular half-life
- The accessibility of a broad wavelength range simply via substitutions and alterations to the TO structure
- The TO1 and TO3 dyes can be used in a 2-color reporter assay system using both the RNA Mango and RNA Peach aptamer systems (Kong et. al. 2021)
TO1-biotin is the standard variety of TO dye for RNA Mango experiments.
The RNA Mango Workflow
Laboratory Results
Mango dye binding to in vitro transcribed RNA Mango-tagged sgRNA. Stable binding and fluorescence even after leaving the tube for 1 month at room temperature.
Transcription reaction were carried out in 300 µL volumes using T7 RNA polymerase (400 U, 50U/µL, applied biological materials), 0.5 µM TO1-3PEG-Biotin (applied biological materials), in 8 mM GTP, 5 mM CTP and ATP, 2 mM UTP, 40 mM TRIS buffer pH 7.9, 2.5 mM spermidine, 26 mM MgCl2, 20 mM KCl, Pyrophosphatase (0.5 U, 0.1 U/µL, ThermoFisher Scientific), and 0.01% Triton X-100. To each sample, either water (Negative), 0.33 µM DNA template (Mango Transcription), or 500 nM final Mango III A10U RNA (Positive) was added. Samples were visualised in a blue light box, movie is played back at 30X speed.
Includes extensive citations list.
Information originally posted on: https://www.abmgood.com/RNA-Mango.html
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