Ammonifex Medium

[所属分类:培养基配方] [发布时间:2019-10-8] [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

Ammonifex Medium
Composition per 1010.0mL:
NaCl..............................................................................................1.0g
Na 2 S·9H 2 O....................................................................................0.5g
K 2 HPO 4 ·3H 2 O ..............................................................................0.3g
KH 2 PO 4 .......................................................................................0.22g
(NH 4 ) 2 SO 4 ..................................................................................0.22g
NaHCO 3 ........................................................................................0.2g
MgSO 4 ·7H 2 O..............................................................................0.09g
CaCl 2 ·2H 2 O ................................................................................0.06g
FeSO 4 ·7H 2 O...............................................................................2.0mg
NiCl 2 ·6H 2 O................................................................................0.2mg
Resazurin...................................................................................0.5mg
KNO 3 solution .........................................................................10.0mL
Selenite/tungstate solution.........................................................3.0mL
Wolfe’s mineral solution............................................................1.0mL
pH 5.4 ± 0.2 at 25°C
Selenite/Tungstate Solution:
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoCl 2 ·6H 2 O..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
KNO 3 Solution:
Composition per 100.0mL:
KNO 3 ..........................................................................................10.0g
Preparation of KNO 3 Solution:  Add KNO 3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except Na 2 S·9H 2 O and KNO 3 solution, to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Add Na 2 S·9H 2 O. Mix thoroughly. Adjust pH to 6.5 with 20% HCl.
Sparge with 100% N 2  for 30 min. Dispense anaerobically into tubes in
10.0mL aliquots. Evacuate headspace under vacuum. Pressurize tubes
to 200 kPa with 80% H 2 + 20% CO 2  gas mixture. Autoclave for 15 min
at 15 psi pressure–121°C. Adjust pH to 5.4. Prior to use, aseptically and
anaerobically add 0.1mL of sterile KNO 3 to each tube.
Use: For the cultivation of Ammonifex species.