Clostridium methylpentosum Medium

[所属分类:培养基配方] [发布时间:2020-5-13] [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 methylpentosum Medium

培养基配方
Composition per liter:
L -Cysteine·HCl ........................................................1.0g
Resazurin ..............................................................1.0mg
Mineral solution A....................................................100.0mL
Mineral solution B....................................................100.0mL
Mineral solution C....................................................100.0mL
NaHCO3  solution.......................................................20.0mL
Rhamnose solution......................................................20.0mL
Vitamin solution.......................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Mineral Solution A:
Composition per 100.0mL:
KH2PO4 .................................................................0.9g
NaCl....................................................................0.9g
NH4Cl ..................................................................0.9g
Preparation of Mineral Solution A:  Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Mineral Solution B:
Composition per 100.0mL:
CaCl2 ·2H2O .........................................................20.0mg
MnCl2 ·4H2O .........................................................20.0mg
CoCl2 ·6H2O...........................................................5.0mg
Preparation of Mineral Solution B:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly.
Mineral solution C:
Composition per 100.0mL
MgSO4 ·7H2O..........................................................20.0mg
FeSO4 ·7H2O...........................................................5.0mg
CuSO4 ·H2O............................................................2.0mg
ZnSO4 ·7H2O...........................................................2.0mg
Preparation of Mineral Solution C:  Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
NaHCO3 Solution:
Composition per 20.0mL:
NaHCO3 ................................................................1.0g
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.
Rhamnose Solution:
Composition per 20.0mL:
Rhamnose ..............................................................2.0g
Preparation of Rhamnose Solution : Add rhamnose to distilled/
deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge
under 100% N2 gas for 3 min. Filter sterilize. Store under N2 gas.
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. Mix thoroughly.
Preparation of Medium: Add components, except NaHCO3 solu-
tion and rhamnose solution, to distilled/deionized water and bring vol-
ume to 960.0mL. Mix thoroughly. Adjust pH to 6.5 with KOH. Sparge
under 100% N2 for 3–4 min. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 20.0mL of sterile NaHCO3
solution and 20.0mL of sterile rhamnose solution. Mix thoroughly.
Aseptically and anaerobically distribute into sterile screw-capped bot-
tles under 100% N2 . Final pH of the medium should be 6.8.
Use:  For the cultivation of Clostridium methylpentosum.