Nonradioactive Detection of Small RNAs Using Digoxigenin-Labeled Probes.

Autor:	Tomassi AH; Instituto de Agrobiotecnología del Litoral (UNL-CONICET-FBCB), Paraje El Pozo, 3000, Santa Fe, Argentina., Gagliardi D; Instituto de Agrobiotecnología del Litoral (UNL-CONICET-FBCB), Paraje El Pozo, 3000, Santa Fe, Argentina., Cambiagno DA; Instituto de Agrobiotecnología del Litoral (UNL-CONICET-FBCB), Paraje El Pozo, 3000, Santa Fe, Argentina., Manavella PA; Instituto de Agrobiotecnología del Litoral (UNL-CONICET-FBCB), Paraje El Pozo, 3000, Santa Fe, Argentina. pablomanavella@ial.santafe-conicet.gov.ar.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2017; Vol. 1640, pp. 199-210.
DOI:	10.1007/978-1-4939-7165-7_14
Abstrakt:	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.
Databáze:	MEDLINE
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