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Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli
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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
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.
