# 1474: PROLIXIBACTER MEDIUM GYSW ## Main sol. 1474 0.54 g NH4Cl 0.14 g KH2PO4 0.2 g MgCl2 x 6 H2O 0.15 g CaCl2 x 2 H2O 10 ml **Trace element solution** 10 ml **Wolin's vitamin solution** 1 ml **Resazurin solution 0.001 g/L** 1.8 g D-Glucose 1 g Yeast extract (Difco) 2.52 g NaHCO3 0.5 g L-Cysteine HCl x H2O 0.5 g Na2S x 9 H2O 1000 ml Distilled water 1. Prepare the liquid medium without Vitamin solution, Cysteine-HCl x H2O and Na2S x 9 H2O. Adjust to pH 7.0 with 1 M NaOH. Dispense the medium into Hungate tubes or serum bottles under gassing with N2:CO2 (80:20 v/v). Sterilize by autoclaving. Before inoculation add from sterile stock solutions Cysteine-HCl x H2O, Vitamin solution (filter sterilized) and Na2S x 9 H2O. ## Trace element solution 12.8 g Nitrilotriacetic acid 1.35 g FeCl3 x 6 H2O 0.1 g MnSO4 x H2O 0.024 g CoCl2 x 6 H2O 0.1 g CaCl2 x 2 H2O 0.1 g ZnCl2 0.025 g CuCl2 x 5 H2O 0.01 g H3BO3 0.024 g Na2MoO4 x 2 H2O 1 g NaCl 0.12 g NiCl2 x 6 H2O 4 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 1000 ml Distilled water 1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH). ## Resazurin solution 0.001 g/L 1 mg Resazurin 1000 ml Distilled water ## Wolin's vitamin solution 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 D-(+)-pantothenate 0.1 mg Vitamin B12 5 mg p-Aminobenzoic acid 5 mg (DL)-alpha-Lipoic acid 1000 ml Distilled water