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