Protocol for making amino acid drop out mixtures for yeast media
This Spreadsheet has a detailed recipe.
Use:
- 2.028 g/L of the complete mixture
- 1.388 g/L of the Dropout (-HULT) = - Histidin Uracile Leucine Trypthophan
- 1.308 g/L of the Dropout (-HULTM) = - Histidin Uracile Leucine Trypthophan Methionine
- 0.640 g/L of the HULT (Histidin, Uracile, Leucine and Tryptophan only)
It can be practical to make stock solutions of amino acids and nucleotides. Some amino acids are not very soluble, using a slightly acid solution usually helps. The data below is from here. The data for Uracile is from here.
| AA | Solubility | final c in medium | times concentrated | Stock soln c | g/ 50 mL | µL/plate (20 mL) |
|---|---|---|---|---|---|---|
| - | g/L | mg/L | stock solution | g/L | ||
| HIS | 41 | 80 | 100 | 8 | 0.4 | 200 |
| URA | 3.6 | 80 | 25 | 2 | 0.1 | 800 |
| LEU | 24 | 400 | 50 | 20 | 1.0 | 400 |
| TRP | 11 | 80 | 100 | 8 | 0.4 | 200 |
| MET | 33 | 80 | 100 | 8 | 0.4 | 200 |
It is practical to make 50 mL of solution for each component as this fit in our plastic syringes (for filtering).
- Add about 20-30 mL of hot dH2O water in a 50 mL falcon tube
- Add the amount specified in the second last column (g/ 50 mL)
- Shake until dissolved.
- Add water to 50 mL, the graduation on the Falcon tube has enough accuracy
- Filter using syringe with 0.2 µM into a sterile Falcon tube
- Store at room temperature, or the solutions may precipitate.