Clostridium thermosulfurogenes Medium

[所属分类:培养基配方] [发布时间:2020-5-18] [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

Clostridium thermosulfurogenes Medium

培养基配方
Composition per 1015.0mL:
Na2HPO4 ·12H2O..........................................................5.3g
NH4Cl ...................................................................1.0g
Yeast extract............................................................1.0g
KH2PO4 ..................................................................0.3g
MgCl2 ·6H2O ............................................................0.2g
FeSO4 ·7H2O............................................................1.5mg
Resazurin ..............................................................1.0mg
Glucose solution ......................................................50.0mL
Trace elements solution................................................10.0mL
Na2S·9H2O solution....................................................10.0mL
Vitamin solution........................................................5.0mL
pH 6.0 ± 0.2 at 25°C
Glucose Solution:
Composition per 50.0mL:
D- Glucose...............................................................5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid...................................................12.5g
NaCl.....................................................................1.0g
FeCl3 ·4H2O.............................................................0.2g
MnCl2 ·4H2O.............................................................0.1g
CaCl2 ·2H2O.............................................................0.1g
ZnCl2 ..................................................................0.02g
CuCl2 ..................................................................0.02g
Na2SeO3 ................................................................0.02g
CoCl2 ·6H2O...........................................................0.017g
H3BO3 ..................................................................0.01g
Na2MoO4 ·2H2O..........................................................0.01g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. AdjustpH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl.......................................................10.0mg
Calcium  DL -pantothenate..............................................5.0mg
Lipoic acid............................................................5.0mg
Nicotinic acid.........................................................5.0mg
p-Aminobenzoic acid....................................................5.0mg
Riboflavin.............................................................5.0mg
Thiamine·HCl .........................................................5.0mg
Biotin ................................................................2.0mg
Folic acid.............................................................2.0mg
Vitamin B 12 ..........................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Adjust pH to 7.0. Mix thor-
oughly. Sparge with 100% N2 .
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.............................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Adjust pH to 7.0 with 1N HCl before use.
Preparation of Medium: Add components, except glucose solution
and Na2S·9H2O solution, and bring volume to 940.0mL. Mix thorough-
ly. Sparge with 80% N2  + 100% CO2 . Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile
glucose solution and 10.0mL of sterile Na2S·9H2O solution. Mix thor-
oughly. Aseptically and anaerobically distribute into tubes or bottles.
Use: For the cultivation and maintenance of Thermoanaerobacterium
thermosulfurigenes.