B693 

MODIFIED AEA SPORULATION MEDIUM BASE

Formula

Ingredients 

gms/lit.

Biopeptone

Yeast extract

Disodium phosphate

Monopotassium phosphate

Ammonium acetate

Magnesiumsulphate 7H2O

10.00

10.00

4.36

0.25

1.50

0.20

Final pH (at 25°C) : 7.8 + 0.2

Directions 

Suspend 26.31 gms. in 1000ml distilled water. Dispense the medium in 15ml amounts in screw capped tubes and sterilize by autoclaving at 15lbs pressure (121°C) for 15 minutes. Cool to 45 – 50°C and add 0.6ml of filter sterilized 10% raffinose and 0.2ml each of sterilie 0.66M sodium carbonate and 0.32% cobalt chloride dropwise to each 15ml base medium in the tubes. Just before use, steam the medium for 10 minutes and after cooling, add 0.2 ml of filter sterilized (freshly prepared) 1.5% sodium ascorbate to each tube of the medium.

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