Erratum: Growth and the Environment of Schizosaccharomyces pombe
Cold Spring Harb Protoc 2016; doi: 10.1101/pdb.top079764
When this article was first published, the recipes for yeast extract with supplements (YES) incorrectly stated that thiamine may be used to repress expression from nmt1 promoters. However, there is sufficient thiamine in the yeast extract to fully repress nmt1 promoters; thiamine is only used in minimal media. The publisher apologizes for this error. The correct versions of the YES recipes are below. In addition, the HTML versions of the recipes and the PDF version of the article have been corrected.
Yeast Extract with Supplements (YES)
| Reagent | Amount to add (1 L) |
|---|---|
| Glucose | 30 g |
| Yeast extract | 5 g |
| Adenine | 0.2 g |
| Uracil | 0.2 g |
| Histidine | 0.2 g |
| Leucine | 0.2 g |
| Lysine | 0.2 g |
| Agar (for solid medium only) | 20 g |
| H2O | to 1 L |
| Prepare 1 L of YES by combining the reagents listed above. Sterilize by autoclaving at 10 psi for 15 min. Store at 4°C. (Note that the yeast extract contains sufficient thiamine to repress nmt1 promoters.) | |
Yeast Extract with Supplements (YES) and 5-Fluoroorotic Acid (5-FOA)
| Reagent | Amount to add (1 L) |
|---|---|
| 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.) | |
Yeast Extract with Supplements (YES) and Phloxin B
| Reagent | Amount to add (1 L) |
|---|---|
| Glucose | 30 g |
| Yeast extract | 5 g |
| Adenine | 0.2 g |
| Uracil | 0.2 g |
| Histidine | 0.2 g |
| Leucine | 0.2 g |
| Lysine | 0.2 g |
| Agar (for solid medium only) | 20 g |
| H2O | to 1 L |
| Prepare 1 L of YES by combining the reagents listed above. Sterilize by autoclaving at 10 psi for 15 min. Add 4 mL of Phloxin B stock solution (5 g/L, filter-sterilized) to 1 L of YES after autoclaving. Store at 4°C. (Note that the yeast extract contains sufficient thiamine to repress nmt1 promoters.) | |
- © 2016 Cold Spring Harbor Laboratory Press
