BA Medium with Cellobiose

[所属分类:培养基配方] [发布时间:2019-11-25] [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

BA Medium with Cellobiose
Composition per liter:
NaHCO3 .....................................................................2.6g
NH4Cl ......................................................................1.0g
Yeast extract..............................................................0.75g
K2HPO4 ·3H2O ..............................................................0.4g
MgCl2 ·6H2O ...............................................................0.1g
NaCl........................................................................0.1g
CaCl2 ·2H2O ..............................................................0.05g
Resazurin .................................................................0.5mg
Cellobiose solution.......................................................50.0mL
Na2S·9H2O solution.......................................................10.0mL
Wolfe’s mineral solution.................................................10.0mL
Wolfe’s vitamin solution.................................................10.0mL
pH 6.9–7.0 at 25°C
Cellobiose Solution:
Composition per 50.0mL:
Cellobiose.................................................................4.0g
Preparation of Cellobiose Solution:  Add cellobiose to distilled/
deionized water and bring volume to 50.0mL. Mix thoroughly. Filter
sterilize.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O................................................................0.25g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O...............................................................3.0g
Nitrilotriacetic acid......................................................1.5g
NaCl.......................................................................1.0g
MnSO4 ·2H2O...............................................................0.5g
CoCl2 ·6H2O ..............................................................0.1g
ZnSO4 ·7H2O...............................................................0.1g
CaCl2 ·2H2O...............................................................0.1g
FeSO4 ·7H2O...............................................................0.1g
NiCl2 ·6H2O.............................................................0.025g
KAl(SO4)2 ·12H2O.........................................................0.02g
CuSO4 ·5H2O..............................................................0.01g
H3BO3 ....................................................................0.01g
Na2MoO4 ·2H2O ...........................................................0.01g
Na2SeO3 ·5H2O............................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl...........................................................10.0mg
p-Aminobenzoic acid........................................................5.0mg
Lipoic acid ...............................................................5.0mg
Nicotinic acid.............................................................5.0mg
Riboflavin ................................................................5.0mg
Thiamine·HCl .............................................................5.0mg
Calcium  DL -pantothenate..................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas mixture. Add components, except cellobiose solu-
tion, Na 2 S·9H 2 O solution, Wolfe’s mineral solution, and Wolfe’s vita-
min solution, to distilled/deionized water and bring volume to
920.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 gas mix-
ture. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and
anaerobically add 50.0mL of sterile cellobiose solution, 10.0mL of
sterile Wolfe’s mineral solutionn, 10.0mL of sterile Wolfe’s vitamin so-
lution, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly.
Aseptically and anaerobically distribute into sterile tubes or bottles.
Use: For the cultivation of Caldicellulosiruptor lactoaceticus.