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Composition | capacity | unit | |||||||||||||||||||||
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Sodium thioglycolate | 0.05 | g | |||||||||||||||||||||
Trypticase peptone (BBL) | 1.0 | g | |||||||||||||||||||||
L-Cysteine·HCl·H2O | 0.1 | g | |||||||||||||||||||||
Heart infusion broth (Difco) | 1.25 | g | |||||||||||||||||||||
Combine the above ingredients and bring to a boil in a round bottom flask while flushing with O2-free N2-H2-CO2 (85:10:5) gas mixture. Stopper anaerobically, wire or seal closed and autoclave at 121C for 20 min. After autoclaving, allow to cool, open without admitting oxygen and while flushing with gas, add the following ingredients which have been filter-sterilized ahead of time: | |||||||||||||||||||||||
Yeast extract (BBL) | 0.25 | g | |||||||||||||||||||||
7.5% NaHCO3 solution | 2.67 | ml | |||||||||||||||||||||
L-Asparagine | 0.25 | g | |||||||||||||||||||||
VFA solution (see below) | |||||||||||||||||||||||
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Distilled water | 94 | ml | |||||||||||||||||||||
Glucose | 0.2 | g | |||||||||||||||||||||
Rabbit serum heat-inactivated at 56C for 1 hr | 2.0 | ml | |||||||||||||||||||||
0.1% Resazurin solution | 0.1 | ml | |||||||||||||||||||||
TPP/VFA mixture (see below) | 2.0 | ml | |||||||||||||||||||||
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** ph : -
** memo : -
** additional actions :