m6A-label-seq : A metabolic labeling protocol to detect transcriptome-wide mRNA N6-methyladenosine (m6A) at base resolution
© 2021 The Author(s)..
We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N6 -methyladenosine (m6A) at base resolution. By feeding cells an analog of methionine, potential mRNA m6A forming sites are replaced with the N6 -allyladenosine (a6A). A mild chemical iodination of a6A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m6A locations could be precisely identified in the high-throughput sequencing. m6A-label-seq provides a strategy to label and identify cellular epitranscriptomic modification sites. For complete details on the use and execution of this profile, please refer to Shu et al., 2020.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:3 |
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| Enthalten in: |
STAR protocols - 3(2022), 1 vom: 18. März, Seite 101096 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Shu, Xiao [VerfasserIn] |
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| Links: |
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| Themen: |
63231-63-0 |
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| Anmerkungen: |
Date Completed 08.04.2022 Date Revised 08.04.2022 published: Electronic-eCollection Citation Status MEDLINE |
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| doi: |
10.1016/j.xpro.2021.101096 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM335942830 |
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| 520 | |a We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N6 -methyladenosine (m6A) at base resolution. By feeding cells an analog of methionine, potential mRNA m6A forming sites are replaced with the N6 -allyladenosine (a6A). A mild chemical iodination of a6A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m6A locations could be precisely identified in the high-throughput sequencing. m6A-label-seq provides a strategy to label and identify cellular epitranscriptomic modification sites. For complete details on the use and execution of this profile, please refer to Shu et al., 2020 | ||
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