Journal
/
/
Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli
DOI:

10:34 min

August 22, 2017

, ,

Chapters

  • 00:05Title
  • 00:37Preparation of Spike-in Cells, Harvesting of Experimental Samples and Addition of Spike-in Cells
  • 02:17RNA Preparation
  • 04:44Gel Electrophoresis, Northern Bloting, and Hybridization
  • 07:43Quantifying tRNA and Charging Levels
  • 08:32Results: Quantification of the Abundance and Charging Levels of tRNAs
  • 09:59Conclusion

Summary

Automatic Translation

Here we present a method for directly measuring transfer RNA charging levels from purified Escherichia coli RNA as well as a way to compare relative levels of transfer RNA, or any other short RNA, across different samples based on the addition of spike-in cells expressing a reference gene.

Related Videos

Read Article