ATCC Medium: 2777 Brockia liithotrophica Medium
Add approximately 0.06 g powdered sulfur per test tube.
Autoclave at 100oC for 30 minutes on 3 consecutive days to sterilize. (Sulfur powder cannot withstand the high heat of the autoclave, so must be sterilized in this manner). On the 3rd day, prepare the medium below:
KCl…………………………………………….……..0.33 g
NH4Cl…………………………………………….….0.33 g
KH2PO4……………………………………………...0.33 g
MgCl2 . 6H2O………………………………………....0.33 g
CaCl2 . 2H2O…………………………………..……..0.33 g
Starch…………………………….…………………5.0 g
Yeast Extract……………………………….……….0.20 g
Resazurin…………………………………………..1 mg
Trace elements (ATCC MD-TMS)………….…….1 ml
DI Water…………………………………………….900 ml
Autoclave at 121oC. After cooling the media, add the following filter sterilized solution:
NaHCO3………………………………………..….. 0.5 g
Na2S . 9H2O………………………………….………0.5 g
Vitamin solution (ATCC MD-VS)…………………1 ml
DI Water……………………………………………..99 ml
Adjust complete medium to pH 6.5 with H2SO4. Prepare under a head space of 80% H2 – 20% CO2.