Geothermobacter Medium (DSMZ Medium 981)

[所属分类:培养基配方] [发布时间:2021-3-11] [发布人: ] [阅读次数:] [返回]

Geothermobacter Medium

(DSMZ Medium 981)

培养基配方

Composition per liter:
NaCl............................................................................................19.0g
MgCl 2 · 6H 2 O..................................................................................9.0g
NaHCO 3 ........................................................................................1.7g
KNO 3 ..........................................................................................1.01g
KCl................................................................................................0.5g
KH 2 PO 4 .......................................................................................0.42g
CaCl 2 ·2H 2 O...................................................................................0.3g
MgSO 4 ·7H 2 O..............................................................................0.15g
Yeast extract..................................................................................0.1g
(NH 4 ) 2 SO 4 .....................................................................................0.1g
NaBr............................................................................................0.05g
SrCl 2 ·6H 2 O..................................................................................0.02g
Resazurin ...................................................................................0.5mg
Vitamin solution.......................................................................10.0mL
NaHCO 3  solution .....................................................................10.0mL
DL -Malic acid solution.............................................................10.0mL
L -Cysteine solution....................................................................5.0mL
Trace elements solution SL-10 ..................................................1.0mL
Selenite/tungstate solution.........................................................1.0mL
pH 6.2 ± 0.2 at 25°C
DL -Malic Acid Solution:
Composition per 10.0mL:
DL -Malic acid..............................................................................1.34g
Preparation of  DL -Malic Acid Solution: Add  D L- malic acid to 10.0mL distilled/deionized water. Mix thoroughly. Sparge with 100%
N 2 . Filter sterilize.
L -Cystine Solution:
Composition per 5.0mL:
L- Cystine-HCl·2H 2 O...................................................................0.05g
NaOH (1N solution)...................................................................5.0mL
Preparation of  L -Cystine Solution: Add  L- cystine to 5.0mL of NaOH solution. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.7g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 20% CO 2 + 80% H 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Trace Elements Solution SL-10:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution SL-10:  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.
Selenite/Tungstate Solution:
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components to distilled/deionized water and bring volume to 970.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................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% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium:  Add components, except bicarbonate, cysteine, vitamins, and malic acid, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Boil for 3 min. Cool to room temperature while sparging with 80% N 2  + 20% CO 2 . Dispense into culture vessels under an atmosphere of 80% N 2  + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anoxically add vitamins, yeast extract, malic acid, and cysteine solution. Adjust pH to 6.2.
Use: For the cultivation of Geothermobacter spp.