Fmoc amino acid (3.5 eq., 175 μmol) and HATU (3.45 eq., 172.5 μmol) are weighed into 10 mL glass vials. Amino acid and coupling reagent are pre-activated by adding 3 mL biotech grade DMF, DIEA (8 eq., 70 μL, 400 μmol), and vortexing. The pre-activated amino acid sits at room temperature for 5 minutes.