Clostridium termitidis Medium

[所属分类:培养基配方] [发布时间:2020-5-14] [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

Clostridium termitidis Medium

培养基配方
Composition per liter:
NaCl........................................................................1.0g
KCl.........................................................................0.5g
Yeast extract...............................................................0.5g
MgCl2 ·6H2O ...............................................................0.4g
NH4Cl ......................................................................0.3g
KH2PO4 .....................................................................0.2g
CaCl2 ·2H2O ..............................................................0.15g
Resazurin..................................................................1.0mg
Trace elements solution SL-10..............................................1.0mL
Cellobiose solution.......................................................50.0mL
NaHCO3  solution..........................................................20.0mL
Na2S·9H2O solution........................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O.................................................................1.5g
CoCl2 ·6H2O..............................................................190.0mg
MnCl2 ·4H2O .............................................................100.0mg
ZnCl2 .....................................................................70.0mg
Na2MoO4 ·2H2O.............................................................36.0mg
NiCl2 ·6H2O...............................................................24.0mg
H3BO3 ......................................................................6.0mg
CuCl2 ·2H2O................................................................2.0mg
HCl (25% solution)..........................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Cellobiose Solution:
Composition per 50.0mL:
D- Cellobiose................................................................5.0g
Preparation of Cellobiose Solution: Add cellobiose to distilled/
deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
under 100% N2 gas for 3 min. Filter sterilize.
NaHCO3 Solution:
Composition per 20.0mL:
NaHCO3 .......................................................................4.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
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. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except cellobiose solu-
tion, NaHCO3  solution, and Na2S·9H2O solution, and bring volume to
920.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2 until pH
reaches below 6.0. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add 50.0mL of sterile cellobiose solu-
tion, 20.0mL of sterile NaHCO3 solution, and 10.0mL of sterile
Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically
distribute into sterile screw-capped bottles under 80% N2 + 20% CO2 .
Use:  For the cultivation and maintenance of Clostridium termitidis.