Protocol To Analyze tRNA and rRNA Processing Using Biotin-Labeled Probes

Authors

Sseu-Pei Hwang
Katherine Barondeau
Catherine Denicourt

Publication Date

6-20-2025

Journal

STAR Protocols

DOI

10.1016/j.xpro.2025.103866

PMID

40448999

PMCID

PMC12166403

PubMedCentral® Posted Date

5-30-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Mature tRNAs and rRNAs are derived from larger precursor transcripts through intricate endo- and exonuclease processing steps. Here, we present a protocol to analyze rRNA and tRNA processing using total RNA extracted from human cells. We describe steps for separating RNA by denaturing electrophoresis, followed by northern blotting. We detail procedures for detecting pre-rRNA intermediates or specific tRNA species using biotin-labeled probes imaged via fluorescence or chemiluminescence. This non-radioactive, high-sensitivity assay enables the investigation of rRNA and tRNA expression dynamics. For complete details on the use and execution of this protocol, please refer to Hwang et al.

Keywords

RNA, Transfer, Biotin, Humans, RNA, Ribosomal, RNA Processing, Post-Transcriptional, Blotting, Northern

Published Open-Access

yes

Recommended Citation

Sseu-Pei Hwang, Katherine Barondeau, and Catherine Denicourt, "Protocol To Analyze tRNA and rRNA Processing Using Biotin-Labeled Probes" (2025). Faculty, Staff and Student Publications. 4338.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/4338