Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines
Summary: Here, we describe a protocol for tRNA identification in the 60S ribosome-nascent peptide complex co-purified with Nuclear Export Mediator Factor (NEMF), a responsible factor for C-terminal alanine and threonine tailing of the nascent peptide. Our protocol is based on regular reverse transcr...
| Päätekijät: | , , |
|---|---|
| Aineistotyyppi: | Artikkeli |
| Kieli: | English |
| Julkaistu: |
Elsevier
2021-09-01
|
| Sarja: | STAR Protocols |
| Aiheet: | |
| Linkit: | http://www.sciencedirect.com/science/article/pii/S2666166721003221 |
| _version_ | 1830343418854768640 |
|---|---|
| author | Tsuyoshi Udagawa Moeka Seki Toshifumi Inada |
| author_facet | Tsuyoshi Udagawa Moeka Seki Toshifumi Inada |
| author_sort | Tsuyoshi Udagawa |
| collection | DOAJ |
| description | Summary: Here, we describe a protocol for tRNA identification in the 60S ribosome-nascent peptide complex co-purified with Nuclear Export Mediator Factor (NEMF), a responsible factor for C-terminal alanine and threonine tailing of the nascent peptide. Our protocol is based on regular reverse transcription followed by quantitative Polymerase chain reaction (PCR). Although this method cannot distinguish between amino acid-charged and uncharged and base-modified and unmodified tRNAs, it is a convenient way to estimate the relative level of tRNA species and thus can be useful for researchers.For complete details on the use and execution of this protocol, please refer to Udagawa et al. (2021). |
| first_indexed | 2024-12-19T22:09:00Z |
| format | Article |
| id | doaj.art-e7ceb7b86cff4beba9b91b6e1cde93db |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-19T22:09:00Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-e7ceb7b86cff4beba9b91b6e1cde93db2022-12-21T20:03:59ZengElsevierSTAR Protocols2666-16672021-09-0123100615Optimized protocol for tRNA identification in the ribosomal complexes from human cell linesTsuyoshi Udagawa0Moeka Seki1Toshifumi Inada2Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 981-8567, Japan; Corresponding authorGraduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 981-8567, JapanGraduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 981-8567, Japan; The institute of Medical Sciences, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; Corresponding authorSummary: Here, we describe a protocol for tRNA identification in the 60S ribosome-nascent peptide complex co-purified with Nuclear Export Mediator Factor (NEMF), a responsible factor for C-terminal alanine and threonine tailing of the nascent peptide. Our protocol is based on regular reverse transcription followed by quantitative Polymerase chain reaction (PCR). Although this method cannot distinguish between amino acid-charged and uncharged and base-modified and unmodified tRNAs, it is a convenient way to estimate the relative level of tRNA species and thus can be useful for researchers.For complete details on the use and execution of this protocol, please refer to Udagawa et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166721003221Cell separation/fractionationMolecular Biology |
| spellingShingle | Tsuyoshi Udagawa Moeka Seki Toshifumi Inada Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines STAR Protocols Cell separation/fractionation Molecular Biology |
| title | Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines |
| title_full | Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines |
| title_fullStr | Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines |
| title_full_unstemmed | Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines |
| title_short | Optimized protocol for tRNA identification in the ribosomal complexes from human cell lines |
| title_sort | optimized protocol for trna identification in the ribosomal complexes from human cell lines |
| topic | Cell separation/fractionation Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721003221 |
| work_keys_str_mv | AT tsuyoshiudagawa optimizedprotocolfortrnaidentificationintheribosomalcomplexesfromhumancelllines AT moekaseki optimizedprotocolfortrnaidentificationintheribosomalcomplexesfromhumancelllines AT toshifumiinada optimizedprotocolfortrnaidentificationintheribosomalcomplexesfromhumancelllines |