520: RUMINOCLOSTRIDIUM CELLULOLYTICUM (CM3) MEDIUM
商品貨號
KDM436
培養(yǎng)基配方
(NH4)2SO4 1.30 g KH2PO4 1.50 g K2HPO4 x 3 H2O 2.90 g FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4) 1.25 ml Trace element solution SL-10 1.00 ml Yeast extract 2.00 g Sodium resazurin (0.1% w/v) 0.50 ml MgCl2 x 6 H2O 0.20 g CaCl2 x 2 H2O 75.00 mg Cellobiose 6.00 g Cellulose, MN 301 (optional) 10.00 g Na2CO3 1.50 g L-Cysteine HCl x H2O 0.50 g Distilled water 1000.00 ml 1. Dissolve ingredients except magnesium chloride, calcium chloride, cellobiose, cysteine and carbonate, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add magnesium chloride, calcium chloride and cellobiose from anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2. Cellobiose has to be sterilized by filtration. Prior to inoculation add cysteine from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH to 7.2. 2. Note: Some strains can be adapted to cellulose as substrate using 10.00 g/l cellulose powder MN 301 (MACHEREY-NAGEL). For DSM 5974, DSM 9801, DSM 17427: Use 5.00 g/l D-glucose as the substrate. Supplement medium after autoclaving with 0.50 g/l Na2S x 9 H2O added from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH of complete medium to 6.8 - 7.0, if necessary. For DSM 27016: Adjust pH of final medium to 8.0. 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.
