ABSTRACT
Agar ..................................................................10.0g Casamino acids ...................................................1.0g Glucose ...............................................................1.0g Tris(hydroxymethyl)aminomethane buffer.........1.0g CaCl2...................................................................0.1g KNO3 ..................................................................0.1g MgSO4·7H2O ......................................................0.1g Sodium glycerophosphate...................................0.1g Thiamine·HCl ..................................................1.0mg Cyanocobalamin .............................................. 1.0µg Trace elements solution HO-LE ..................... 1.0mL
pH 7.5 ± 0.2 at 25°C
Trace Elements Solution HO-LE: Composition per liter: H3BO3 ...............................................................2.85g MnCl2·4H2O .......................................................1.8g Sodium tartrate..................................................1.77g FeSO4·7H2O......................................................1.36g CoCl2·6H2O ......................................................0.04g CuCl2·2H 2O ...................................................0.027g Na2MoO4·2H2O ..............................................0.025g ZnCl2.................................................................0.02g
Preparation of Trace Elements Solution HOLE: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except trace elements solution HO-LE, to distilled/ deionized water and bring volume to 999.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-121°C. Cool to 45°–50°C. Aseptically add 1.0mL of trace elements solution HO-LE. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
