Sulfolobus Plates (YS or T) (download as file)
(ala Zillig)
for 1 liter (revised September 2007)
1. Dissolve in approx. 800 ml distilled water:
a. 3.0 grams Ammonium sulfate: (NH4)2SO4
b. 0.7 grams Glycine
c. 0.5 grams Potassium hydrogen phosphate (potassium phosphate, dibasic): K2HPO4 d. 0.1 grams Potassium chloride: KCl
2. Carbon source: YS:
a. 1.0 gram Yeast extract b. 2.0 grams Sucrose
Tryptone (T):
3. Trace elements (solutions are on shelf above the scale):
a. 200 μl 1% Ferrous sulfate: FeSO4
b. 244 μl 1% Sodium borate: Na2B4O7 • 10 H2O c. 90 μl Manganese chloride: MnCl2 • 4 H2O
d. 11 μl Zinc sulfate: ZnSO4 • 7 H2O
e. 2.5 μl Cupric sulfate: CuSO4 • 5 H2O
f. 1.5 μl Sodium molybdate: Na2MoO4 • 2 H2O g. 1.5 μl Vanadyl sulfate: VOSO4 • 5 H2O
h. 0.5 μl Cobalt chloride: CoCl2 • 6 H2O
i. 0.5 μl Nickel sulfate: NiSO4 • 6 H2O
4. Adjust pH to 3.0 – 3.2 with 50% sulfuric acid (approx. 300 μl)
5. Add distilled water to make 1 liter
6. Dissolve 10 grams of Gelrite in 1 liter of media by boiling (add Gelrite slowly to gently boiling media) or autoclaving for 20 minutes on the liquid/fluid cycle.
7. Add 5 ml of 1M magnesium chloride / 0.3 M calcium nitrate solution (pre-made) after the Gelrite has completely dissolved (adding the Mg/Ca solution before the Gelrite has dissolved will prevent the Gelrite from completely dissolving, leading to plates that melt in the incubator)
8. Pour the plates in the laminar flow hood.