Final pH: 7.2 - 7.4 Final volume: 1003 ml Solution A 942.00 ml Solution B 30.00 ml Solution C 20.00 ml Solution D 1.00 ml Solution E 10.00 ml Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic, distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are prepared under 100% N2 gas and sterilized by filtration. Solution E is autoclaved under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust the pH of the complete medium to 7.2 – 7.4, if necessary. For DSM 5848, DSM 9537: Replace glucose with 2.50 g/l of Na2-succinate and 0.50 g/l yeast extract as substrates. For DSM 6233: Replace glucose with 0.50 g/l of pyrogallol as the substrate. When growth appears feed once more with 0.50 g/l pyrogallol. For DSM 12881, DSM 19012, DSM 25728: Supplement medium with 0.50 g/l yeast extract. For DSM 15285, DSM 19306: Replace glucose with 10.00 g/l Proteose peptone (BD Difco) as substrate. Adjust pH of complete medium to 6.5 - 7.0. For DSM 17953, DSM 19335: Replace glucose with 2.50 g/l maltose and 2.00 g/l yeast extract as substrates. For DSM 106009: Use 0.30 g/l L-Cysteine-HCl x H2O and 1.50 g/l acetaldehyde as substrates. Acetaldehyde should be added in aliquots of 0.20 g/l at regular intervals until full growth is achieved. For DSM 108915: Supplement medium with 3.2 g Na2-fumarate, 2.0 g Yeast extract, and 0.3 g L-Cysteine HCl x H2O. Omit D-Glucose and Na2S x 9 H2O. Adjust pH of final medium to 6.5. Solution A KH2PO4 0.20 g NH4Cl 0.25 g NaCl 20.00 g MgCl2 x 6 H2O 3.00 g
KCl 0.50 g CaCl2 x 2 H2O 0.15 g Trace element solution SL-10 1.00 ml Selenite-tungstate solution 1.00 ml Sodium resazurin (0.1% w/v) 0.50 ml Distilled water 940.00 ml Solution B Na2CO3 1.50 g Distilled water 30.00 ml Solution C D-Glucose 2.00 g Distilled water 20.00 ml Solution D Seven vitamins solution 1.00 ml Solution E Na2S x 9 H2O 0.30 g Distilled water 10.00 ml Trace element solution SL-10 (from medium 320) HCl (25%) 10.00 ml FeCl2 x 4 H2O 1.50 g ZnCl2 70.00 mg MnCl2 x 4 H2O 100.00 mg H3BO3 6.00 mg CoCl2 x 6 H2O 190.00 mg CuCl2 x 2 H2O 2.00 mg NiCl2 x 6 H2O 24.00 mg Na2MoO4 x 2 H2O 36.00 mg Distilled water 990.00 ml First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. Seven vitamins solution (from medium 503) Vitamin B12 100.00 mg p-Aminobenzoic acid 80.00 mg D-(+)-biotin 20.00 mg Nicotinic acid 200.00 mg Calcium pantothenate 100.00 mg Pyridoxine hydrochloride 300.00 mg Thiamine-HCl x 2 H2O 200.00 mg Distilled water 1000.00 ml Selenite-tungstate solution (from medium 385) NaOH 0.50 g Na2SeO3 x 5 H2O 3.00 mg Na2WO4 x 2 H2O 4.00 mg Distilled water 1000.00 ml
