Optimized protocol for quantifying 5' UTR-mediated translation initiation in S. cerevisiae using direct analysis of ribosome targeting.
STAR Protoc
; 3(4): 101862, 2022 12 16.
Article
em En
| MEDLINE
| ID: mdl-36595943
ABSTRACT
Direct analysis of ribosome targeting (DART) allows investigators to measure the translation initiation potential of thousands of RNAs in parallel. Here, we describe an optimized protocol for generating active translation extract from S. cerevisiae, followed by in vitro translation, purification of ribosome-bound RNAs, and subsequent library preparation and sequencing. This protocol can be applied to a variety of cell types and will enable high-throughput interrogation of translational determinants. For complete details on the use and execution of this protocol, please refer to Niederer et al. (2022).1.
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Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
/
Biossíntese de Proteínas
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article