Long undecoded transcript isoform (LUTI) detection in meiotic budding yeast by direct RNA and transcript leader sequencing

STAR Protocols

Folkert J. van Werven¹  Minghao Chia²  Amy Tresenrider³  Elçin Ünal⁴ 

[1] Corresponding author;Department of Genome Sciences, Foege Hall, University of Washington, Seattle, WA 98105, USA;Department of Molecular and Cell Biology, Barker Hall, University of California, Berkeley, Berkeley, CA 94720, USA;Genome Institute of Singapore, 60 Biopolis Street, Genome, 02-01, Singapore 138672, Singapore;

关键词: Bioinformatics;    Sequence analysis;    Genetics;    Genomics;    Sequencing;    Model Organisms;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: LUTIs (Long Undecoded Transcript Isoforms) are 5′-extended and poorly translated mRNAs that can downregulate transcription from promoters more proximal to a gene’s coding sequence (CDS). In this protocol, polyA RNA is extracted from budding yeast cells undergoing highly synchronized meiosis. Using a combination of long-read direct RNA sequencing and transcript leader sequencing (TL-seq), meiosis-specific LUTIs are systematically identified. Following identification, TL-seq is used to quantify the abundance of both LUTI and the more canonical gene-proximal (PROX) transcripts.For complete details on the use and execution of this protocol, please refer to Tresenrider et al. (2021).

【 授权许可】

Unknown