FISH-Flow to quantify nascent and mature ribosomal RNA in mouse and human cells.
STAR Protoc
; 4(3): 102463, 2023 Sep 15.
Article
en En
| MEDLINE
| ID: mdl-37481729
ABSTRACT
FISH-Flow (fluorescence in situ hybridization-flow cytometry) involves hybridizing fluorescent oligos to RNA and quantifying fluorescence at a single-cell level using flow cytometry. Here, we present a FISH-Flow protocol to quantify nascent 47S and mature 18S and 28S rRNAs in mouse and human cells, including rRNA quantification across cell cycle stages using DNA staining. We describe steps for cell preparation, hybridization of fluorescent probes against rRNA, and DNA staining. We then detail procedures for flow cytometry and data analysis. For complete details on the use and execution of this protocol, please refer to Antony et al. (2022).1.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ARN
/
ARN Ribosómico
Límite:
Animals
/
Humans
Idioma:
En
Revista:
STAR Protoc
Año:
2023
Tipo del documento:
Article
País de afiliación:
Estados Unidos