ATCC medium: 2223 Anaerobic serine/arginine medium
NH4Cl ..........................................0.3 g
K2HPO4 .........................................0.2 g
KH2PO4 .........................................0.3 g
MgCl2 . 6H2O ...................................0.4 g
CaCl2 . 2H2O ...................................0.15 g
KCl............................................0.5 g
NaCl...........................................1.0 g
SL-10 Trace Element Solution (see below).......1.0 ml
Selenite/Tungstate Solution (see below)........1.0 ml
Yeast extract..................................2.0 g
Serine*........................................1.05 g
Resazurin......................................0.5 mg
Distilled water..............................940.0 ml
* Arginine-HCl, 1.74 g, may be used in place of the serine Prepare medium anaerobically under 80% N2, 20% CO2.
Autoclave at 121C for 15 minutes. Cool under the same gas mixture.
After cooling, add the following filter-sterilized ingredients:
Na2S . 9H2O Solution (see below)
NaHCO3 Solution (see below)
Wolfe's Vitamin Solution (see below)..........10.0 ml
Adjust final pH of the completed medium to 7.2.
Dispense anaerobically as desired.
SL-10 Trace Element Solution:
HCl (25%)...................10.0 ml
FeCl2 . 4H2O .................1.5 g
ZnCl2 .......................70.0 mg
MnCl2 . 4H2O ...............100.0 mg
H3BO3 ........................6.0 mg
CoCl2 . 6H2O ...............190.0 mg
CuCl2 . 2H2O .................2.0 mg
NiCl2 . 6H2O ................24.0 mg
Na2MoO4 . 2H2O...............36.0 mg
Distilled water............990.0 ml
Dissolve FeCl2 in the HCl, dilute with water, add and dissolve the other salts; adjust pH to 6.0 with NaOH, and fill to 1.0 L with distilled water.
Selenite/Tungstate Solution:
NaOH.........................0.5 g
Na2SeO3 . 5H2O................3.0 mg
Na2WO4 . 2H2O.................4.0 mg
Distilled water..............1.0 L
Na2S . 9H2O Solution:
Na2S . 9H2O ..................0.6 g
Distilled water.............10.0 ml
NaHCO3 Solution:
NaHCO3 .......................5.0 g
Distilled water.............50.0 ml
Wolfe's Vitamin Solution:
Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).
Biotin.......................2.0 mg
Folic acid...................2.0 mg
Pyridoxine hydrochloride....10.0 mg
Thiamine . HCl...............5.0 mg
Riboflavin...................5.0 mg
Nicotinic acid...............5.0 mg
Calcium D-(+)-pantothenate...5.0 mg
Vitamin B12..................0.1 mg
p-Aminobenzoic acid..........5.0 mg
Thioctic acid................5.0 mg
Distilled water..............1.0 L