| Glucose |
60 g |
| Yeast extract |
10 g |
| Adenine |
0.4 g |
| Uracil |
0.1 g |
| Histidine |
0.4 g |
| Leucine |
0.4 g |
| Lysine |
0.4 g |
| Agar (for solid medium only) |
40 g |
| H2O
|
to 1 L |
| Prepare 1 L of 2× YES by combining the reagents listed above. Sterilize by autoclaving at 10 psi for 15 min. Dissolve 2 mg
of 5-FOA (Sigma-Aldrich F5013) in 1 mL of sterile milliQ H2O. Mix 1:1 with 55°C 2× YES using a sterile stir bar. (5-FOA from Sigma-Aldrich is very expensive; cheaper reagents from other
suppliers do not work as well. Calculate the quantities that you will require to make up the precise number of plates you
need for your selection.) Store at 4°C. (Note that the yeast extract contains sufficient thiamine to repress nmt1 promoters.)
|
