Clostridium thermolacticum 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 thermolacticum Medium

培养基配方
Composition per 1020.0mL:
KHCO3 ...........................................................................4.5g
NaCl............................................................................2.25g
Sucrose..........................................................................2.0g
Yeast extract....................................................................2.0g
MgSO4 ·7H2O.....................................................................0.5g
NH4Cl ...........................................................................0.5g
K2HPO4 ........................................................................0.348g
CaCl2 ·2H2O....................................................................0.25g
KH2PO4 ........................................................................0.227g
FeSO4 ·7H2O....................................................................2.0mg
Resazurin ......................................................................1.0mg
L -Cysteine·HCl·H2O solution .................................................10.0mL
Na2S·9H2O solution.............................................................10.0mL
Wolfe’s vitamin solution.......................................................10.0mL
Trace elements solution SL-6 ....................................................3.0mL
pH 7.0–7.2 at 25°C
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl.................................................................10.0mg
p-Aminobenzoic acid..............................................................5.0mg
Calcium pantothenate ............................................................5.0mg
Nicotinic acid...................................................................5.0mg
Riboflavin ......................................................................5.0mg
Thiamine·HCl ...................................................................5.0mg
Thioctic acid ...................................................................5.0mg
Biotin ..........................................................................2.0mg
Folic acid.......................................................................2.0mg
Cyanocobalamin ...............................................................100.0μg
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly.
Trace Elements Solution SL-6:
Composition per liter:
H3BO3 .........................................................................0.3g
CoCl2 ·6H2O ..................................................................0.2g
ZnSO4 ·7H2O...................................................................0.1g
MnCl2 ·4H2O .................................................................0.03g
Na2MoO4 ·H2O.................................................................0.03g
NiCl2 ·6H2O..................................................................0.02g
CuCl2 ·2H2O..................................................................0.01g
Preparation of Trace Elements Solution SL-6:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 3.4.
L - Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O..........................................................0.3g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -
cysteine·HCl·H2O to distilled/deionized water and bring volume to
10.0mL. Gas tubes under 100% N2 and tightly seal. Autoclave for 15
min at 15 psi pressure–121°C. Use freshly prepared solution.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.....................................................................0.3g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Gas tube under
100% N2 and tightly seal. Autoclave for 15 min at 15 psi pressure–
121°C. Use freshly prepared solution.
Preparation of Medium:  Prepare anaerobically under 80% N2 +
20% CO2 . Add components, except  L -cysteine·HCl·H2O solution and
Na2S·9H2O solution, to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Distribute into tubes using anaerobic techniques.
Autoclave for 15 min at 15 psi pressure–121°C. Prior to inoculation of
cultures, inject 0.1mL of sterile  L -cysteine·HCl·H2O solution and 0.1mL
of sterile Na2S·9H2O solution per 10.0mL of medium.
Use:  For the cultivation and maintenance of Clostridium thermolacti-
cum.