Medium data

Search for medium no. = [1248].

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

1248 MINERAL SALTS MEDIUM WITH TCE

NaCl	`1.0`	g
MgCl₂·6H₂O	`0.5`	g
KH₂PO₄	`0.2`	g
NH₄Cl	`0.3`	g
KCl	`0.3`	g
CaCl₂·2H₂O	`0.015`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Selenite tungstate solution (see Medium No. 431)	`1.0`	ml
Sodium formate	`0.34`	g
Sodium acetate	`0.41`	g
Resazurin	`0.25`	mg
Distilled water	`1.0`	L

Mix components thoroughly, bring to a boil and cool down under a N₂-CO₂ (4:1, v/v) gas mixture. Add the following solutions:

8% NaHCO₃ solution	`30.0`	ml
5% L-Cysteine·HCl·H₂O solution	`6.0`	ml
5% Na₂S·9H₂O solution	`10.0`	ml

Adjust pH of the medium to 7.0-7.2, if necessary. Dispense the medium into culture vessels (e.g., 100 ml in 120 ml-serum bottles) under the same gas mixture and seal with butyl rubber stoppers or Teflon-lined stoppers. Then, anaerobically add per liter the following solution:

Tetrachloroethene 60.0 μl

Dissolve tetrachloroethene with ultrasound. Autoclave in a closed basket. Do not remove the culture vessels from autoclave before temperature of the media has reached below 70C.
Finally, add per liter the following solution from an anaerobic stock (filter-sterilized):

Trace vitamins (see Medium No. 197) 10.0 ml