Yeast tRNA extract was acquired from Sigma Aldrich (Roche 10109495001) and desalted using piperidine-based mobile phases on a Dionex RSLCnano Ultimate 3000 as described previously. A short desalting column (5 cm by 100 µm ID fused silica with a homemade Kasil frit) was packed in-house using Waters XBridge Phenyl (3.5 µm, 130 Å) pH-resistant stationary phase. For each run, 0.5 µL of a 10 µM solution of yeast tRNA extract was injected and desalted with an isocratic gradient of 80% B (A: water, 10 mM piperidine; B: acetonitrile, 10 mM piperidine) at a flow rate of 1 µL/min. This nanoflow desalting approach resulted in a 10-minute run, enabling high-throughput analysis.