ATCC medium: 1995 Thiobacillus caldus medium
Solution A:
(NH4)2SO4.....................3.0 g
Na2SO4 . 10H2O................3.2 g
KCl..........................0.1 g
K2HPO4 ......................50.0 mg
MgSO4 . 7H2O .................0.5 g
Distilled water............460.0 ml
Adjust solution A to pH 1.75 with H2SO4.
Autoclave at 121C for 15 minutes. Maintain above 60C (see below).
Solution B:
Ca(NO3)2 . 4H2O..............10.0 mg
FeCl3 . 6H2O ................11.0 mg
CuSO4 . 5H2O .................0.5 mg
H3BO3 ........................2.0 mg
MnSO4 . H2O ..................2.0 mg
Na2MoO4 . 2H2O................0.8 mg
CoCl2 . 6H2O .................0.6 mg
ZnSO4 . 7H2O .................0.9 mg
Distilled water.............10.0 ml
Filter-sterilize solution B.
Solution C:
Glucose......................0.45 g
Distilled water.............10.0 ml
Filter-sterilize solution C.
Solution D:
Sodium tetrathionate.........0.77 g
Distilled water.............20.0 ml
Filter-sterilize solution D.
Solution E:
Phytagel (Sigma P-8169).....15.0 g
Distilled water............500.0 ml
Autoclave solution E at 121C for 15 minutes.
Maintain above 60C (see below). Maintain solutions A and E above 60C when removed from autoclave to retard rapid gelling of Phytagel. Aseptically combine solutions A through E and dispense into appropriate vessels. Complete medium will have a final pH of 2.5.
For liquid medium, omit solution E and prepare solution A in 960 ml water. Adjust solution A to pH 2.5.
For chemolithotrophic growth, omit glucose.