15n2 uracil

S. cerevisiae BY4741 was grown in ¹³C labeled Yeast OD 2 (+ 10 g/L ¹³C₆ Glucose (99% ATOM) (Euriso-top, Saclay, France)) rich medium (Silantes, Munich, Germany) for SILIS preparation or in YNB minimal medium for NAIL-MS experiments. 10x YNB (Carl Roth GmbH, Karlsruhe, Germany) was prepared according to manufacturer's manual. 1x YNB medium was supplemented with 10 g/L glucose, 0.02 g/L uracil, a mix of needed unlabeled aminoacids (final concentration: 0.02 g/L arginine, 0.02 g/L histidine, 0.06 g/L leucine, 0.03 g/L lysine, 0.05 g/L phenylalanine, 0.4 g/L serine, 0.2 g/L threonine, 0.04 g/L tryptophane, 0.03 g/L tyrosine and 0.15 g/L valine) and additional needed aminoacids used for labeling (final concentration: 0.1 g/L aspartic acid, 0.1 g/L glutamine and 0.02 g/L methionine). Depending on the desired labeling ¹⁵N₂-uracil, L-aspartic-¹⁵N acid, L-glutamin-amide-¹⁵N, L-methionine-methyl-D₃ or their unlabeled isotopomers were used. ¹⁵N₂-uracil (98% ATOM ¹⁵N) was supplied by Euriso-top. All unlabeled aminoacids and L-aspartic-¹⁵N acid (98% ATOM ¹⁵N), L-glutamin-amide-¹⁵N (98 ATOM% ¹⁵N) and L-methionine-methyl-D3 (98% ATOM D₃) were supplied by Sigma-Aldrich, Munich, Germany.