Brackish Prosthecomicrobium Medium

[所属分类:培养基配方] [发布时间:2020-1-3] [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

Brackish Prosthecomicrobium Medium

培养基配方
Composition per liter:
Agar ........................................................15.0g
Peptone......................................................0.25g
Yeast extract................................................0.25g
Glucose .....................................................0.25g
Artificial seawater........................................250.0mL
Modified Hutner’s basal salts .............................20.0mL
Vitamin solution............................................10.0mL
pH 7.2 ± 0.2 at 25°C
Artificial Seawater:
Composition per liter:
NaCl........................................................23.477g
MgCl2 .......................................................4.981g
Na2SO4 ......................................................3.917g
CaCl2 .......................................................1.102g
KCl..........................................................0.664g
NaHCO3 ......................................................0.192g
KBr..........................................................0.096g
H3BO3 .......................................................0.026g
SrCl2 .......................................................0.024g
NaF...........................................................3.0mg
Preparation of Artificial Seawater:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly.
Modified Hutner’s Basal Salts:
Composition per liter:
MgSO4· 7H2O ..................................................29.7g
Nitrilotriacetic acid..........................................10.0g
CaCl2 ·2H2O...................................................3.34g
FeSO4 ·7H2O...................................................0.1g
(NH4)2MoO4 ....................................................9.25mg
Metals “44”..................................................50.0mL
Preparation of Modified Hutner’s Basal Salts:  Dissolve the
nitrilotriacetic acid first and neutralize the solution with KOH. Add the
other ingredients and readjust the pH with KOH and/or H 2 SO 4  to 7.2.
There may be a slight precipitate. Store at 5°C.
Metals “44”:
Composition per 100.0mL:
ZnSO4 ·7H2O...................................................1.1g
FeSO4 ·7H2O...................................................0.5g
EDTA..........................................................0.25g
MnSO4 ·7H2O.................................................0.154g
CuSO4 ·5H2O..................................................0.04g
Co(NO3)2 ·6H2O..............................................0.025g
Na2B4O7 ·10H2O..............................................0.018g
Preparation of Metals “44”:  Add components to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Add aseptically to sterile basal
medium.
Vitamin Solution:
Composition per liter:
Thiamine·HCl.................................................5.0mg
D- Calcium pantothenate.......................................5.0mg
Riboflavin....................................................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. Filter ster-
ilize and add aseptically to sterile basal medium.
Preparation of Medium: Add a few drops of H 2 SO 4 to the distilled
water to retard precipitation of the metal salts. Add components, except
Metals “44” and vitamin solution, to 250.0mL of artificial seawater and
720.0mL of distilled/deionized water. Adjust pH to 7.2. Distribute into
tubes or flasks. Sterilize by autoclaving for 15 min at 15 psi pressure–
121°C. Aseptically add Metals “44” and vitamin solution. Mix thor-
oughly. Aseptically distribute into sterile tubes or flasks.
Use:  For the cultivation of Prosthecomicrobium litoralum.