Medium data

Search for medium no. = [462].

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

462 SALTWATER MEDIUM WITH LACTATE

NaCl	`20.0`	g
Na₂SO₄	`4.0`	g
MgCl₂·6H₂O	`3.0`	g
CaCl₂·2H₂O	`0.15`	g
NH₄Cl	`0.75`	g
KH₂PO₄	`0.6`	g
KBr	`0.09`	g
KCl	`0.5`	g
NaHCO₃	`2.5`	g
Trace element solution (see Medium No. 439)	`1.0`	ml
Vitamin solution (see Medium No. 403)	`1.0`	ml
Thiamine solution (see Medium No. 403)	`1.0`	ml
Vitamin B₁₂ solution (see Medium No. 403)	`1.0`	ml
Riboflavin (see below)	`1.0`	ml
Selenite-tungstate solution (see Medium No. 431)	`1.0`	ml
Sodium lactate	`2.25`	g
Na₂S·9H₂O	`0.3`	g
Resazurin	`1.0`	mg

Dissolve components except NaHCO₃, Vitamin solution, Thiamine solution, Vitamin B₁₂ solution, sodium lactate and Na₂S·9H₂O in 900 ml distilled water and adjust pH to 7.0. Bring to a boil and cool down under a N₂-CO₂ (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 9 ml of the medium in Hungate tubes) under a N₂-CO₂ (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Separately autoclave 0.2 M sodium lactate under a N₂ atmosphere. After cooling, add 1/9 volume of the sodium lactate solution, and filter-sterilized 8% NaHCO₃ solution, Vitamin solution, Thiamine solution and Vitamin B₁₂ solution to the medium. Readjust pH to 7.0, if necessary. Prior to use, reduce the medium with Na₂S·9H₂O (3% solution, autoclaved and stocked under N₂).
Riboflavin solution:

Riboflavin	`2.5`	mg
Distilled water	`100.0`	ml