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Main sol. J144

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Internal Solution-ID: S3788

Equipment needed: Autoclave Anaerobic workbench Filter
1 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:
Compound Amount Unit Conc. [g/L] Conc. [mM]
Heart Infusion Broth
(BD-Difco) 		1.25 g 12.376 -
Trypticase peptone
(BD-BBL) 		1.00 g 9.901 -
Yeast extract
(BD-BBL) 		0.25 g 2.475 -
Sodium thioglycolate 0.05 g 0.495 4.339
L-Cysteine HCl x H2O 0.10 g 0.99 -
L-Asparagine 0.25 g 2.475 18.735
Glucose 0.20 g 1.98 10.991
Resazurin
(0.1%) 		0.10 ml - -
Distilled water 94.00 ml - -
NaHCO3
(7.5%) 		2.67 ml - -
TPP/VFA mixture 2.00 ml - -
Rabbit serum
(heat-inactivated at 56C for 1 hr) 		2.00 ml - -
J144 MODIFIED NOS MEDIUM