Northern Blotting: Protocols for Radioactive and Nonradioactive Detection of RNA.

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Contributed Indexing:	Keywords: 32P-labelled oligonucleotide; Digoxigenin; Long noncoding RNA; Nonradioactive Northern blotting; Northern blotting; Total RNA extraction; Xrn1-sensitive lncRNA; Yeast; mRNA decay
Substance Nomenclature:	63231-63-0 (RNA) 0 (Phosphorus Radioisotopes) NQ1SX9LNAU (Digoxigenin) 0 (RNA, Fungal) 0 (Oligonucleotide Probes)
Entry Date(s):	Date Created: 20241113 Date Completed: 20241113 Latest Revision: 20241113
Update Code:	20241115
DOI:	10.1007/978-1-0716-4176-7_2
PMID:	39535701
Autor:	Wery M; ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France. maxime.wery@curie.fr., Foretek D; ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France., Andjus S; ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France., Verdys P; ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France., Morillon A; ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France. antonin.morillon@curie.fr.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2025; Vol. 2863, pp. 13-28.
DOI:	10.1007/978-1-0716-4176-7_2
Abstrakt:	Northern blotting is a common technique in RNA biology, allowing to detect and quantify RNAs of interest following separation by gel electrophoresis, transfer to a membrane, and hybridization of specific anti-complementary labelled probes. In this chapter, we describe our protocol for efficient RNA extraction from yeast, separation on agarose gel, and capillary transfer to a membrane. We provide two different methods for strand-specific detection of several types of RNAs using oligonucleotide probes, the first using radioactive 32 P-labelled probes, the second based on nonradioactive digoxigenin-labelled probes. (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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