ABSTRACT
Agar ..................................................................18.0g Na2HPO4 .........................................................0.732g KH2PO4...........................................................0.466g NaCl ..................................................................0.29g Sodium propionate..............................................0.2g MgSO4·7H2O ......................................................0.1g CaCO3 ...............................................................0.02g KNO3 ................................................................0.01g FeSO4·7H2O.....................................................0.2mg ZnSO4·7H2O ..................................................0.18mg MnSO4·4H2O .................................................0.02mg Cycloheximide solution ................................ 10.0mL Thiamine·HCl solution.................................. 10.0mL
pH 7.0 ± 0.2 at 25°C
Cycloheximide Solution: Composition per 10.0mL: Cycloheximide ..................................................0.05g
Preparation of Cycloheximide Solution: Add cycloheximide to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Thiamine·HCl Solution: Composition per 10.0mL: Thiamine·HCl ..................................................4.0mg
Preparation of Thiamine·HCl Solution: Add thiamine·HCl to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except cycloheximide solution and thiamine·HCl solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure121°C. Cool to 45°–50°C. Aseptically add 10.0mL of sterile cycloheximide solution and 10.0mL of thiamine·HCl solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
