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  3. Protocol for rapid tRNA enrichment and chemiluminescent northern blot detection of tRNA and tRNA-derived fragments

Protocol for rapid tRNA enrichment and chemiluminescent northern blot detection of tRNA and tRNA-derived fragments

  • STAR Protoc. 2026 Mar 20;7(1):104357. doi: 10.1016/j.xpro.2026.104357.
Pavlina Gregorova 1,  Minna-Maria K Heinonen 2,  Milla M Laarne 2,  L Peter Sarin 3
Affiliations

Affiliations

  • 1 RNAcious Laboratory, Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, Viikinkaari 9B, 00790 Helsinki, Finland. Electronic address: [email protected].
  • 2 RNAcious Laboratory, Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, Viikinkaari 9B, 00790 Helsinki, Finland.
  • 3 RNAcious Laboratory, Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, Viikinkaari 9B, 00790 Helsinki, Finland. Electronic address: [email protected].
PMID: 41653438 DOI: 10.1016/j.xpro.2026.104357
Abstract

Transfer RNA (tRNA), its post-transcriptional modifications, and tRNA-derived fragments (tRFs) play essential roles in cellular processes and gene regulation. Here, we present a fast and efficient tRNA enrichment protocol using silica spin columns. To analyze tRNA and tRFs, we describe steps for DNA probe labeling, tRNA separation on polyacrylamide (PAA) gels, and RNA transfer, UV crosslinking, and non-radioactive northern blotting. Additionally, this protocol describes the use of chemical affinity modifiers, enabling the detection of chemical modifications in specific tRNA isoacceptors.

Keywords

Cell Biology; Molecular Biology; Molecular/Chemical Probes.

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