Medium data

Search for medium no. = [448].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

448   KA22 MEDIUM

Sea salts (Sigma)  30.0     g
Mg(NO3)·6H2O  2.54    g
KH2PO4  0.3     g
NaHCO3  1.0     g
MES  1.95    g
Sulfur (powder)  12.0     g
Trace minerals (see Medium No. 151)  10.0     ml
Trace vitamins (see Medium No. 197)  0.5     ml
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients except sulfur, NaHCO3 and Na2S·9H2O, adjust pH to 6.5 and autoclave. Filter-sterilize 8% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S·9H2O solution under a N2 atmosphere. Aseptically distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml culture medium in 120 ml serum bottle containing 0.24 g sulfur) under a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S·9H2O solution and readjust pH to 6.5, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).

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