Fig. 5.
In-vivo aminoacylation assays for mitochondrial tRNA. Equal amounts (2 μg) of total mitochondrial RNA purified from various cell lines under acid conditions were treated with electrophoresis at 4°C through an acid (pH 5.1) 10% polyacrylamide/7 mol/l urea gel, electroblotted onto a positively charged nylon membrane, and hybridized with digoxigenin (DIG)-labeled oligonucleotide probes specific for mitochondrial tRNAThr. Samples were also deacylated (DA) by heating 10 min at 60°C at pH 8.3. The blots were then stripped and rehybridized with DIG-labeled probes for tRNALeu(CUN), tRNASer(AGY), tRNALys, and tRNAGly, respectively.