Enriching s4 U-RNA Using Methane Thiosulfonate (MTS) Chemistry.
Curr Protoc Chem Biol
; 8(4): 234-250, 2016 Dec 07.
Article
em En
| MEDLINE
| ID: mdl-27925666
ABSTRACT
Metabolic labeling of cellular RNA is a useful approach to study RNA biology. 4-Thiouridine (s4 U) is a convenient nucleoside for metabolic labeling because it is cell permeable and is incorporated into newly transcribed RNA, and the sulfur moiety provides a handle for biochemical purification. However, a critical step in the purification of s4 U-RNA is the efficiency of the chemistry used to enrich s4 U-RNA. Here, we present a protocol for s4 U-RNA enrichment that includes efficient and reversible covalent chemistry to biotinylate s4 U-RNA using the activated disulfide methane thiosulfonate conjugated to biotin (MTS-biotin), followed by enrichment on streptavidin beads. The efficiency of this chemistry reduces enrichment bias and requires less starting material, thereby expanding the utility of s4 U to study RNA biology. © 2016 by John Wiley & Sons, Inc.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Tiouridina
/
RNA
/
Mesilatos
Limite:
Humans
Idioma:
En
Revista:
Curr Protoc Chem Biol
Ano de publicação:
2016
Tipo de documento:
Article