Anaerobic Citrate Medium

[所属分类:培养基配方] [发布时间:2019-10-14] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Anaerobic Citrate Medium
Composition per liter:
Ferric citrate................................................................................17.0g
Sodium acetate..............................................................................6.8g
NaHCO 3 ........................................................................................2.5g
NH 4 Cl ...........................................................................................1.5g
NaH 2 PO 4 ·H 2 O...............................................................................0.6g
KCl................................................................................................0.1g
Trace elements solution ...........................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
Na 2 WO 4 ...................................................................................25.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
Wolfe’s mineral solution...............................................................1.0L
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
CaCl 2 .............................................................................................0.1g
CoCl 2 ·6H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
CuSO 4 ·5H 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 approximately 500.0mL of water and adjust to pH 6.5 with
KOH to dissolve the compound. Bring volume to 1.0L with remaining
water and add remaining compounds one at a time.
Preparation of Trace Elements Solution:  Combine compo-
nents. Mix thoroughly.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add ferric citrate to 500.0mL of boiling distilled/de-
ionized water. Mix thoroughly. Cool to room temperature while sparg-
ing with 80% N 2 + 20% CO 2 . Add remaining components. Add
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 7.0 with NaOH. Continue sparging with 80% N 2 + 20%
CO 2 . Anaerobically distribute into tubes. Autoclave for 15 min at 15
psi pressure–121°C.
Use: For the cultivation of Geobacter metallireducens.