1) Create a Dropout Powder Stock
| 1 | L-Arginine monohydrochloride | 0.02 g |
| 2 | L-Aspartic acid monosodium hydrate | 0.1 g |
| 3 | L-Glutamic acid potassium salt monohydrate | 0.1 g |
| 4 | L-Isoleucine | 0.03 g |
| 5 | L-Lysine monochloride | 0.03 g |
| 6 | L-Phenylalanine | 0.05 g |
| 7 | L-Serine | 0.375 g |
| 8 | L-Threonine | 0.2 g |
| 9 | L-Tyrosine | 0.03 g |
| 10 | L-Valine | 0.15 g |
| Total | 1.085 g | |
Dissolved in 1 liter of H20, the amounts of amino acids and nucleobases above result in 1x solutions.
No: Ade, His, Leu, Met, Trp, or Ura.
Shake up powder stock before using.
Store the powder stock at 4°C.
2) Add for 1L of SC medium
– 10 g Succinic Acid (C4H6O4)
– 6 g Sodium Hydroxide (NaOH)
– 5 g Ammonium Sulfate (NH4)2SO4
– 1.7 g YNB (w/o amino acids or ammonium sulfate) (If YNB contains ammonium sulfate, then add 5 g + 1.7 g = 6.7 g of YNB)
– 1.085 g Dropout Powder Stock (SC-AHLMUW)
To add particular amino acids or nucleobases, pick the ones needed from the table of amino acids and nucleobases (1x) below:
| 11 | Adenine hemisulfate | 0.04 g/l |
| 12 | L-Histidine monohydrochloride monohydrate | 0.02 g/l |
| 13 | L-Leucine | 0.12 g/l |
| 14 | L-Methionine | 0.02 g/l |
| 15 | L-Tryptophan | 0.08 g/l |
| 16 | Uracil | 0.02 g/l |
Example: To make plates without Methionine only, add to 1.085 g of the Dropout Powder Stock: 0.04 g Ade, 0.02 g Ura, 0.02 g His, 0.12 g Leu, and 0.08 g Trp.
Once everything has been added, mix well and bring H2O up to 900mL and put in a 1L bottle.
3) For plates
Add 20 g Agar
Autoclave (35 min)
After autoclave:
Stir until cool enough to touch
Add 100 mL of 10x sugar solution (20% Glucose, 30% Galactose, or 30% Raffinose stock)
Pour plates
Author: Roxane Dervey
updated: October 2020