Magnetic Spirillum Growth Medium, Revised (MSGM, Revised)

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Magnetic Spirillum Growth Medium, Revised
(MSGM, Revised)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Agar ..............................................................................................1.3g
KH 2 PO 4 .......................................................................................0.68g
Tartaric acid ................................................................................0.37g
Succinic acid...............................................................................0.37g
NaNO 3 .........................................................................................0.12g
Sodium acetate............................................................................0.05g
Ascorbic acid ............................................................................0.035g
Wolfe’s vitamin solution..........................................................10.0mL
Wolfe’s mineral solution............................................................5.0mL
Ferric quinate solution...............................................................2.0mL
Resazurin (0.1% solution)........................................................0.45mL
pH 6.75 ± 0.2 at 25°C
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl ............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium pantothenate ................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid ..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Cyanocobalamin .....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution : Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitriloacetic acid...........................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O ..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Preparation of Wolfe’s Mineral Solution:  Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L.
Ferric Quinate Solution:
Composition per 100.0mL:
FeCl 3 ...........................................................................................0.27g
Quinic acid..................................................................................0.19g
Preparation of Ferric Quinate Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium:  To 1.0L of distilled/deionized water add
components in the following order: Wolfe’s vitamin solution, Wolfe’s min-
eral solution, ferric quinate solution, resazurin, KH 2 PO 4 , NaNO 3 , ascorbic
acid, tartaric acid, succinic acid, sodium acetate, and agar. Mix thoroughly
after each addition. Adjust pH to 6.75 with NaOH. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically distribute into sterile screw-capped
tubes. Fill tubes to capacity with medium. Use a heavy inoculum in each
tube and do not introduce a headspace of air. Screw down caps tightly.
Use:  For the cultivation and maintenance of Aquaspirillum magnetot-
acticum.