ATCC medium: 1653 Revised magnetic Spirillum growth medium (MSGM)
Distilled water...........................1.0 L
Wolfe's Vitamin Solution (see below)......10.0 ml
Wolfe's Mineral Solution (see below)......5.0 ml
0.01 M Ferric Quinate (see below).........2.0 ml
0.1% Resazurin............................0.45 ml
KH2PO4 ....................................0.68 g
NaNO3 .....................................0.12 g
Ascorbic acid.............................0.035 g
Tartaric acid.............................0.37 g
Succinic acid.............................0.37 g
Sodium acetate............................0.05 g
Agar (for semi-solid medium)..............1.3 g
Add components in the order given with stirring. Adjust to pH 6.75 with NaOH.
Liquid Medium:
Autoclave medium at 121C for 15 minutes. Aseptically fill screw-capped containers to full capacity with sterile medium. Inoculate heavily leaving no headspace of air, and screw down closures tightly.
Semi-Solid Medium: Dispense 12 ml per 16 X 150 mm screw-cap tube.
Autoclave at 121C for 15 minutes.
0.01 M Ferric Quinate:
FeCl3 .....................................0.27 g
Quinic Acid (Sigma Q-0500)................0.19 g
Distilled water...........................100.0 ml
Dissolve and autoclave at 121C for 15 minutes.
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
2 Distilled water...........................1.0 L