Medium 155: DOUBLE LAYER AGAR FOR PHAGE PROPAGATION
First layer:
Nutrient agar 28 g
Yeast extract powder 3 g
Peptone 5 g
Distilled water 1000 ml
Adjust pH to 7.4 +/- 0.2.
Sterilized at 121°C for 15 min.
Second layer (per 1 plate):
0.7% meat peptone agar (45°C) 2.5 ml
bacterial culture* 0.1 ml
phage suspenzion 0.1 ml
*Add sterile solution of CaCl2 (stock solution
0.02M) to final concentration 0.002M.
0.7% meat peptone agar:
Nutrient broth 13 g
Yeast extract powder 3 g
Peptone 5 g
Agar no. 1 7 g
Distilled water 1000 ml
Adjust pH to 7.4 +/- 0.2.
Sterilized at 121°C for 15 min.