Abstract
Small RNAs have been traditionally detected and quantified using small RNA blots, a modified Northern blot technique. The small RNAs are size-fractionated from the rest of the cellular RNA molecules by polyacrylamide gel electrophoresis and transferred by blotting onto a positively charged membrane. A radiolabeled probe was then traditionally used to detect a specific small RNA in the cellular pool. Small RNA blotting is a relatively simple, inexpensive approach to visualize small RNAs without artifacts. However, the radioactive labeling of the probe is sometimes an impediment, especially due to the requirement of specialized facilities. Here we describe a sensitive and simple method to detect and quantify small RNAs using digoxigenin-based nonradioactive RNA blots.
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Acknowledgments
The laboratory of P.A.M. is funded by the Human Frontier Science Program, the Max Planck Society, the International Centre for Genetic Engineering and Biotechnology, and the Agencia Nacional de Promoción Científica y Tecnológica, Argentina. P.A.M. is a member of Consejo Nacional de Investigaciones Científicas y Técnicas. A.H.T. and D.G are fellows of the same institution.
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Tomassi, A.H., Gagliardi, D., Cambiagno, D.A., Manavella, P.A. (2017). Nonradioactive Detection of Small RNAs Using Digoxigenin-Labeled Probes. In: Carbonell, A. (eds) Plant Argonaute Proteins. Methods in Molecular Biology, vol 1640. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7165-7_14
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DOI: https://doi.org/10.1007/978-1-4939-7165-7_14
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