ATCC Medium: 1470 Modified Leptospira Medium
Peptone (BD 211677)………………………….0.3 g
Beef Extract……………………………………..0.2 g
NaCl……………………………………………..0.5 g
Agar……………………………………………...1.5 g
DI Water…………………………………………900 ml
Adjust basal medium for a final pH of 7.4. Autoclave at 121C.
Cool to 50 C and add aseptically:
Sterile Rabbit Serum…………………………….100 ml
*0.05% Hemin (see below)………….…………..2.5 ml
Adjust final pH to 7.3 +/- 0.1. Inactivate entire medium for 1 hr at 56C the day following its preparation. 0.05% Hemin Dissolve Hemin in 0.5-1.0 ml of 1N NaOH.
Then add distilled water to 100 ml. Sterilize by autoclave at 121C or if preferred, by filter sterilization.