Clostridium thermoaceticum Medium

[所属分类:培养基配方] [发布时间:2020-5-15] [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 thermoaceticum Medium

培养基配方
Composition per 1010.0mL:
Solution A...............................................................100.0mL
Solution B...............................................................600.0mL
Solution C...............................................................300.0mL
Solution D................................................................10.0mL
pH 6.9 ± 0.2 at 25°C
Solution A:
Composition per 100.0mL:
Glucose ...................................................................18.0g
Preparation of Solution A: Add glucose to distilled/deionized wa-
ter and bring volume to 100.0mL. Mix thoroughly. Sparge with 100%
N2  for 5–10 min. Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 600.0mL:
Pancreatic digest of casein..................................................5.0g
Yeast extract................................................................5.0g
Pyruvic acid ................................................................1.8g
(NH4)2SO4 ...................................................................1.0g
MgSO4 ·7H2O................................................................0.25g
Fe (NH4)2 (SO4)2 ·6H2O.....................................................0.04g
Co(NO3)2 ·6H2O ............................................................0.03g
Na2WO4 ·2H2O...............................................................3.3mg
Na2MoO4 ·2H2O..............................................................2.4mg
ZnCl2 ......................................................................1.4mg
Resazurin...................................................................1.0mg
Na2SeO3 ·5H2O..............................................................0.3mg
NiCl2 ·6H2O................................................................0.2mg
Preparation of Solution B:  Add components to distilled/deionized
water and bring volume to 600.0mL. Mix thoroughly. Sparge with
100% CO2  for 5–10 min. Autoclave for 15 min at 15 psi pressure–
121°C.
Solution C:
Composition per 300.0mL:
NaHCO3 ....................................................................16.8g
K2HPO4 .....................................................................7.0g
KH2PO4 .....................................................................5.5g
Preparation of Solution C:  Add components to distilled/deionized
water and bring volume to 300.0mL. Mix thoroughly. Sparge with
100% CO2  for 5–10 min. Autoclave for 15 min at 15 psi pressure–
121°C.
Solution D:
Composition per 10.0mL:
L -Cysteine·HCl solution (5%) ..............................................5.0mL
Na2S·9H2O solution (5%).....................................................5.0mL
Preparation of Solution D:  Combine 5.0mL of  L -cysteine·HCl so-
lution and 5.0mL of Na2S·9H2O solution. Mix thoroughly. Sparge with
100% N2  for 5–10 min. Autoclave for 15 min at 15 psi pressure–121°C.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl ............................................................0.5g
Preparation of  L -Cysteine·HCl Solution:  Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 80% N2 + 20% CO2 for 15 min at 15 psi pressure–
121°C.
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.
Preparation of Medium: Aseptically and anaerobically combine
100.0mL of sterile solution A, 600.0mL of sterile solution B, 300.0mL
of sterile solution C, and 10.0mL of sterile solution D. Mix thoroughly.
Aseptically and anaerobically distribute into tubes or bottles.
Use:  For the cultivation and maintenance of Clostridium thermoaceti-
cum.