Medium data

Search for medium no. = [1103].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1103 ANAEROLINEA MEDIUM C

Solution A:

KH₂PO₄	`0.14`	g
MgCl₂·6H₂O	`0.2`	g
CaCl₂·2H₂O	`0.15`	g
NH₄Cl	`0.54`	g
Yeast extract (BD-Difco)	`2.3`	g
Trace element solution SL-11 (see below)	`1.0`	ml
Selenite tungstate solution (see Medium No. 431)	`0.5`	ml
Trace vitamins (see Medium No. 197)	`10.0`	ml
NaHCO₃	`2.5`	g
Resazurin	`1.0`	mg
Distilled water	`900.0`	ml

Mix components except NaHCO₃ and trace vitamins, and adjust pH to 6.5. Bring to a boil for 5-10 sec, then, cool down under a stream of N₂-CO₂ (80:20, v/v) and add NaHCO₃ to the solution. Dispense Solution A into culture vessels under a stream of N₂-CO₂ (80:20, v/v), seal with butyl rubber stopper and autoclave. After autoclaving, stand the medium overnight. Add trace vitamins from a sterile anaerobic stock (filter-sterilized).
Solution B:

Glucose	`2.2`	g
Distilled water	`100.0`	ml

Adjust pH to 7.0 and filter-sterilize. Keep under a N₂ atmosphere.
To complete the medium, aseptically and anaerobically add 0.1 volume of Solution B to 0.9 volume of Solution A (e.g., 18 ml Solution A in 60 ml serum bottles and 2 ml Solution B). Prior to inoculation, add per liter the following solutions:

5% Na₂S·9H₂O solution	`6.0`	ml
5% L-Cysteine·HCl·H₂O solution	`6.0`	ml

Trace element solution SL-11:

EDTA·2Na	`5.2`	g
FeCl₂·4H₂O	`1.5`	g
ZnCl₂	`70.0`	mg
MnCl₂·4H₂O	`100.0`	mg
H₃BO₃	`6.0`	g
CoCl₂·6H₂O	`190.0`	mg
CuCl₂·2H₂O	`2.0`	mg
NiCl₂·6H₂O	`24.0`	mg
Na₂MoO₄·2H₂O	`36.0`	mg
Distilled water	`1.0`	L

Adjust pH to 6.0.