FB Medium (DSMZ Medium 980)

[所属分类:培养基配方] [发布时间:2020-7-13] [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

FB Medium
(DSMZ Medium 980)

培养基配方
Composition per liter:
NH4Cl ....................................................................0.54g
MgCl2 ·6H2O...............................................................0.2g
CaCl2 ·2H2O..............................................................0.15g
KH2PO4 ...................................................................0.14g
Resazurin ................................................................0.5mg
NaHCO3 solution...........................................................10.0mL
Yeast extract solution....................................................10.0mL
Na-crotonate solution.....................................................10.0mL
Cysteine solution.........................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Vitamin solution..........................................................10.0mL
Trace elements solution SL-9 ..............................................1.0mL
Selenite tungstate solution................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Na-Crotonate Solution:
Composition per 10.0mL:
Na-crotonate................................................................0.86g
Preparation of Na-Crotonate Solution:  Add Na-crotonate to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.

Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O.......................................................0.25g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H2O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Trace Elements Solution SL-9:
Composition per liter:
MgSO4 ·7H2O................................................................3.0g
Nitrilotriacetic acid.......................................................1.5g
NaCl........................................................................1.0g
MnSO4 ·2H2O................................................................0.5g
CoSO4 ·7H2O...............................................................0.18g
ZnSO4 ·7H2O...............................................................0.18g
CaCl2 ·2H2O ...............................................................0.1g
FeSO4 ·7H2O................................................................0.1g
NiCl2 ·6H2O..............................................................0.025g
KAl(SO4)2 ·12H2O..........................................................0.02g
H3BO3 .....................................................................0.01g
Na2MoO4 ·4H2O.............................................................0.01g
CuSO4 ·5H2O...............................................................0.01g
Na2SeO3 ·5H2O.............................................................0.3mg
Preparation of  Trace Elements Solution SL-9: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl...........................................................10.0mg
Thiamine-HCl·2H2O........................................................5.0mg
Riboflavin................................................................5.0mg
Nicotinic acid............................................................5.0mg
D -Ca-pantothenate .......................................................5.0mg
p-Aminobenzoic acid.......................................................5.0mg
Lipoic acid...............................................................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. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.25g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .....................................................................2.5g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract...............................................................0.2g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Preparation of Medium: Add components, except Na-crotonate
solution, yeast extract solution, cysteine solution, vitamin solution,
NaHCO3 solution, and Na2S·9H2O solution, to distilled/deionized wa-
ter and bring volume to 940.0mL. Mix thoroughly. Gently heat and
bring to boiling. Boil for 3 min. Cool to 25°C while sparging with 80%
N2 + 20% CO2 . Distribute to anaerobe tubes or bottles under 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptical-
ly and anaerobically add per liter of medium, 10.0mL sterile cysteine
solution, 10.0mL sterile Na-crotonate solution, 10.0mL sterile vitamin
solution, 10.0mL sterile yeast extract solution, 10.0mL sterile NaHCO3
solution, and 10.0mL sterile Na2S·9H2O solution. Mix thoroughly. The
final pH should be 7.0.
Use:  For the cultivation of Sporotomaculum syntrophicum.