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An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity
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Biochemistry
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JoVE Journal Biochemistry
An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

DOI:
10.3791/58100-v

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07:46 min

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October 08, 2018

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, , , , ,
1Department of Biology,Ball State University, 2Department of Genome Sciences,University of Washington, 3Department of Molecular, Cellular, and Developmental Biology,University of Michigan

Chapters

  • 00:04Title
  • 00:39Preparing a 20% Polyacrylamide Denaturing Gel
  • 02:17RNA Isopentenylation Assay
  • 05:14Results: Detection of Isopentenylation by Mod5
  • 06:42Conclusion

Summary

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Here, we describe a protocol for the biochemical characterization of the yeast RNA-modifying enzyme, Mod5, and discuss how this protocol could be applied to other RNA-modifying enzymes.

Tags

Keywords: TRNA-Isopentenyl TransferaseIn Vitro AssayRNA ModificationBiochemical CharacterizationRecombinant EnzymesRNA Modifying EnzymesAcrylamide GelUrea GelTBE BufferRNA Isopentenylation AssayDMAPPMod52-mercaptoethanolRNA Precipitation

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