Protocol to analyze tRNA and rRNA processing using biotin-labeled probes
Summary: 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 elect...
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| Language: | English |
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Elsevier
2025-06-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002722 |
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| author | Sseu-Pei Hwang Katherine Barondeau Catherine Denicourt |
| author_facet | Sseu-Pei Hwang Katherine Barondeau Catherine Denicourt |
| author_sort | Sseu-Pei Hwang |
| collection | DOAJ |
| description | Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-9a4e61b800e5413aacedb7fa7f6e1db2 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-9a4e61b800e5413aacedb7fa7f6e1db22025-08-20T02:17:28ZengElsevierSTAR Protocols2666-16672025-06-016210386610.1016/j.xpro.2025.103866Protocol to analyze tRNA and rRNA processing using biotin-labeled probesSseu-Pei Hwang0Katherine Barondeau1Catherine Denicourt2Department of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center, Houston, TX 77030, USA; The University of Texas MD Anderson Cancer Center UTHealth Houston Graduate School of Biomedical Sciences, Houston, TX 77030, USA; Corresponding authorDepartment of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center, Houston, TX 77030, USA; Corresponding authorDepartment of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center, Houston, TX 77030, USA; The University of Texas MD Anderson Cancer Center UTHealth Houston Graduate School of Biomedical Sciences, Houston, TX 77030, USA; Corresponding authorSummary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002722Cell BiologyGeneticsMolecular BiologyMolecular/Chemical Probes |
| spellingShingle | Sseu-Pei Hwang Katherine Barondeau Catherine Denicourt Protocol to analyze tRNA and rRNA processing using biotin-labeled probes STAR Protocols Cell Biology Genetics Molecular Biology Molecular/Chemical Probes |
| title | Protocol to analyze tRNA and rRNA processing using biotin-labeled probes |
| title_full | Protocol to analyze tRNA and rRNA processing using biotin-labeled probes |
| title_fullStr | Protocol to analyze tRNA and rRNA processing using biotin-labeled probes |
| title_full_unstemmed | Protocol to analyze tRNA and rRNA processing using biotin-labeled probes |
| title_short | Protocol to analyze tRNA and rRNA processing using biotin-labeled probes |
| title_sort | protocol to analyze trna and rrna processing using biotin labeled probes |
| topic | Cell Biology Genetics Molecular Biology Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002722 |
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