Coprothermobacter proteolyticus Medium

[所属分类:培养基配方] [发布时间:2020-5-26] [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

Coprothermobacter proteolyticus Medium

培养基配方
Composition per 1168.1mL:
Yeast extract.........................................................2.0g
Trypticase™...........................................................2.0g
NaOH solution.........................................................1.0L
Gelatin solution...................................................113.0mL
Na2S solution.......................................................22.6mL
Solution A..........................................................10.0mL
Mineral salts solution..............................................10.0mL
Solution B...........................................................2.0mL
Resazurin solution ..................................................0.5mL
NaOH Solution:
Composition per liter:
NaOH..................................................................4.0g
Preparation of NaOH Solution: Add NaOH to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly.
Gelatin Solution:
Composition per 100.0mL:
Gelatin................................................................3.0g
Preparation of Gelatin Solution: Gently heat 100.0mL of dis-
tilled/deionized water to 80°C. Sparge with 100% N 2 for 15 min. Add
the gelatin. Mix thoroughly. Sparge with 100% N 2 for 10 min. Auto-
clave for 15 min at 15 psi pressure–121°C.
Na2S Solution:
Na2S...................................................................2.5g
Distilled water.......................................................100ml
Preparation of Na2S Solution: Gently heat 100.0mL of distilled/
deionized water to 100°C. Boil for 5 min. Sparge with 100% N 2 for 15
min. Add the Na2S. Mix thoroughly. Sparge with 100% N 2 for 10 min.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution A:
Composition per liter:
NH4Cl .................................................................100.0g
MgCl2 ·H2O ...........................................................100.0g
CaCl2 ·2H2O ...........................................................40.0g
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 4 with
HCl.
Mineral Salts Solution:
Composition per liter:
EDTA·2H2O..............................................................0.5g
CoCl2 ·H2O............................................................0.15g
MnCl2 ·4H2O ...........................................................0.1g
FeSO4 ·7H2O............................................................0.1g
ZnCl2 ..................................................................0.1g
AlCl3 ·H2O.............................................................40mg
Na2WO4 ·2H2O...........................................................30mg
CuCl2 ·2H2O............................................................20mg
NiSO4 ·H2O.............................................................20mg
H2SeO3 .................................................................10mg
H3BO4 ..................................................................10mg
NaMoO4 ·2H2O...........................................................10mg
Preparation of Mineral Salts Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Ad-
just pH to 3 with HCl.
Solution B:
Composition per liter:
K2HPO4 ·3H2O.........................................................200.0g
Preparation of Solution B: Add K2HPO4 ·3H2O to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly.
Resazurin Solution:
Composition per 100.0mL:
Resazurin ..............................................................0.2g
Preparation of Resazurin Solution: Add resazurin to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly.
Preparation of Medium:  Sparge 1.0L of NaOH solution with
100% CO2 for 30 min. Add 2.0g of yeast extract and 2.0g of Trypti-
case™. Mix thoroughly. Add 10.0mL of solution A, 2.0mL of solution
B, 0.5mL of resazurin solution, and 10.0mL of mineral salts solution
with pipets which have been flushed a few times with 100% N 2 . Mix
thoroughly. Anaerobically distribute 9.0mL volumes into anaerobic
tubes fitted with butyl rubber stoppers. Autoclave for 15 min at 15 psi
pressure–121°C. One hour prior to inoculation, add 1.0mL of sterile
gelatin solution and 0.2mL of sterile Na2S solution to each 9.0mL of
medium.
Use: For the cultivation of Coprothermobacter proteolyticus.