Figure 1D - tRNA acylation levels of exponentially growing MG1655 and MG1655relA::HTF measured by Northern blotting analysis

tRNA acylation levels of exponentially growing MG1655 and MG1655relA::HTF measured by northern blotting analysis. Total tRNA was extracted from cells growing in MOPS minimal medium with 0.2% glucose as the carbon source before and after isoleucine starvation. A fraction of the purified tRNA was deacylated (by 100 mM Tris-HCL [pH 9]) and charged and uncharged samples were separated by urea PAGE and transferred to a membrane. tRNAileTUV, tRNAasnUTVW and tRNAtrpT were detected by hybridization with radiolabeled tRNA-specific DNA oligonucleotide probes.