# J977: VNG MEDIUM ## Main sol. J977 100 ml **10 x CMD stock solution** 10 ml **Trace vitamins** 0.027 g NH4Cl 3.57 g HEPES 790 ml Distilled water 1. Mix components thoroughly, adjust pH to 8.1 with NaOH or HCl. Distribute the medium into culture vessels (e.g., 9 ml in 25 ml serum bottles/Balch tubes), seal with butyl rubber stoppers and autoclave. After cooling, add the following solution (per 900 ml, autoclaved): 100 ml Glucose (w/v, 20%) ## 10 x CMD stock solution 1 g Nitrilotriacetic acid 1 ml **5 x CMD micronutrient solution** 4.8 mg FeCl3 x 6 H2O 0.6 g CaSO4 x 2 H2O 1 g MgSO4 x 7 H2O 0.8 g NaCl 1.03 g KNO3 6.89 g NaNO3 1.11 g NaH2PO4 1000 ml Distilled water 1. Do not use if exceeds 90 days shelf life. ## 5 x CMD micronutrient solution 100 ml Distilled water 0.25 ml H2SO4 (6.25 N) 1.14 g MnSO4 x n H2O 0.25 g ZnSO4 x 7 H2O 0.25 g H3BO3 8.5 mg CuCl2 x 2 H2O 12.5 mg Na2MoO4 x 2 H2O 22.5 mg CoCl2 x 6 H2O ## Trace vitamins 2 mg Biotin 2 mg Folic acid 10 mg Pyridoxine hydrochloride 5 mg Thiamine HCl 5 mg Riboflavin 5 mg Nicotinic acid 5 mg Calcium pantothenate 0.1 mg Vitamin B12 5 mg p-Aminobenzoic acid 5 mg Lipoic acid 1000 ml Distilled water