Preparation of Spike-in Cells, Harvesting of Experimental Samples and Addition of Spike-in Cells
2:17
RNA Preparation
4:44
Gel Electrophoresis, Northern Bloting, and Hybridization
7:43
Quantifying tRNA and Charging Levels
8:32
Results: Quantification of the Abundance and Charging Levels of tRNAs
9:59
Conclusion
副本
The overall goal of this experiment is to quantify the abundance and charging levels of transfer RNAs in Escherichia coli. This method can help answer key questions in cell biology such as, how tRNA charging levels changes at different growth stat
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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.