TO1-3PEG-Desthiobiotin Fluorophore
| Cat. No. | G956 | ||
| Name | TO1-3PEG-Desthiobiotin Fluorophore | ||
| Unit | 0.5 mg/ml (500 µl) | ||
| Category | RNA Tracking (RNA Mango) | ||
| Description |
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 apatmer (Fluorescent enhancement FE=1,100).
TO1-biotin is the standard variety of TO dye for RNA Mango experiments; other dye variants are available. Learn more about the RNA Mango technology here.
Watch RNA Mango in Action 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 visualized in a blue light box, movie is played back at 30X speed.
|
||
| Storage Condition |
Stored at -20°C and protect from light. |
||
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G956 |
| Can I use water with my fluorophore dyes? | |
|
We strongly recommend to use DMF, DMSO, 10% Acetonitrile, or MeOH-CH₂ Cl₂ for stability. Do no store in water as it may break down the dye.
|
| Which aptamers should I use for each RNA Mango fluorophore? | |
|
Use our RNA Mango Aptamer Systems chart to help you select the appropriate aptamer and fluorophore combination for your research needs.
|
| Any tips for using RNA Mango aptamers in cellular transcripts? | |
|
We have an aptamer insertion guideline for cellular transcript, which can be found here.
|
| What is the difference between TO1-3PEG-Biotin (G955) and TO1-3PEG-Desthiobiotin (G956)? | |
|
TO1-3PEG-Biotin (G955) and TO1-3PEG-Desthiobiotin (G956) are quite similar in terms of binding and fluorescence. They are distinct in how they interact with streptavidin magnetic beads. Specifically, desthiobiotin can be displaced from them by the addition of free biotin while TO1-3PEG-Biotin cannot.
|
- Autour, A., C Y Jeng, S., D Cawte, A., Abdolahzadeh, A., Galli, A., Panchapakesan, S. S. S., Rueda, D., Ryckelynck, M., & Unrau, P. J. (2018). Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells. Nature communications, 9(1), 656. https://doi.org/10.1038/s41467-018-02993-8
- Trachman, R. J., & Ferré-D'Amaré, A. R. (2019). Tracking RNA with light: selection, structure, and design of fluorescence turn-on RNA aptamers. Quarterly reviews of biophysics, 52, e8. https://doi.org/10.1017/S0033583519000064
- Trachman, R. J., 3rd, Autour, A., Jeng, S. C. Y., Abdolahzadeh, A., Andreoni, A., Cojocaru, R., Garipov, R., Dolgosheina, E. V., Knutson, J. R., Ryckelynck, M., Unrau, P. J., & Ferré-D'Amaré, A. R. (2019). Structure and functional reselection of the Mango-III fluorogenic RNA aptamer. Nature chemical biology, 15(5), 472–479. https://doi.org/10.1038/s41589-019-0267-9
- Kong, K. Y. S., Jeng, S. C. Y., Rayyan, B., & Unrau, P. J. (2021). RNA Peach and Mango: Orthogonal two-color fluorogenic aptamers distinguish nearly identical ligands. RNA (New York, N.Y.), 27(5), 604–615. Advance online publication. https://doi.org/10.1261/rna.078493.120
- Cawte, A. D., Unrau, P. J., & Rueda, D. S. (2020). Live cell imaging of single RNA molecules with fluorogenic Mango II arrays. Nature communications, 11(1), 1283. https://doi.org/10.1038/s41467-020-14932-7
- Panchapakesan, S. S. S., Ferguson, M. L., Hayden, E. J., Chen, X., Hoskins, A. A., & Unrau, P. J. (2017). Ribonucleoprotein purification and characterization using RNA Mango. RNA (New York, N.Y.), 23(10), 1592–1599. https://doi.org/10.1261/rna.062166.117
- Mitra, J., & Ha, T. (2019). Nanomechanics and co-transcriptional folding of Spinach and Mango. Nature communications, 10(1), 4318. https://doi.org/10.1038/s41467-019-12299-y
- Shi, J., Gao, X., Tian, T., Yu, Z., Gao, B., Wen, A., You, L., Chang, S., Zhang, X., Zhang, Y., & Feng, Y. (2019). Structural basis of Q-dependent transcription antitermination. Nature communications, 10(1), 2925. https://doi.org/10.1038/s41467-019-10958-8
- Fang, C., Philips, S. J., Wu, X., Chen, K., Shi, J., Shen, L., Xu, J., Feng, Y., O'Halloran, T. V., & Zhang, Y. (2021). CueR activates transcription through a DNA distortion mechanism. Nature chemical biology, 17(1), 57–64. https://doi.org/10.1038/s41589-020-00653-x