MAB1 Medium

[所属分类:培养基配方] [发布时间:2021-5-11] [发布人:网站管理员2] [阅读次数:] [返回]

MAB1 Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1003.0mL:

NaCl............................................................................................20.0g
MgCl 2 ·6H 2 O .................................................................................3.0g
Na 2 SO 4 ..........................................................................................3.0g
KCl................................................................................................0.5g
NH 4 Cl .........................................................................................0.25g
Yeast extract..................................................................................0.2g
KH 2 PO 4 .........................................................................................0.2g
Sodium benzoate.........................................................................0.15g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin...................................................................................1.0mg
Wolfe’s vitamin solution..........................................................20.0mL
NaHCO 3  solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Na 2 SeO 3 /Na 2 WO 4 solution........................................................1.0mL
Sodium dithionite solution.........................................................1.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
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.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize. Sparge with 80% N 2  + 20% CO 2 .
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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.
Na 2 SeO 3 /Na 2 WO 4 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 Na 2 SeO 3 /Na 2 WO 4 Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Sodium Dithionite Solution:
Composition per 10.0mL:
Sodium dithioninium ....................................................................0.2g
Preparation of Sodium Dithionite Solution:  Add sodium dithi-
oninium to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Prepare and dis-
pense under 80% N 2 + 20% CO 2 . Add FeCl 2 ·4H 2 O to 10.0mL of HCl
solution. Mix thoroughly. Add distilled/deionized water and bring vol-
ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with
80% N 2  + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare medium and dispense under 80%
N 2 + 20% CO 2 . Add components, except Wolfe’s vitamin solution,
NaHCO 3 solution, sodium dithionite solution, Na 2 S·9H 2 O solution,
Na 2 SeO 3 /Na 2 WO 4 solution, and trace elements solution SL-10, to dis-
tilled/deionized water and bring volume to 960.0mL. Mix thoroughly.
Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15
min at 15 psi pressure–121°C. Aseptically and anaerobically add
20.0mL of Wolfe’s vitamin solution, 10.0mL of sterile NaHCO 3 solu-
tion, 10.0mL of sterile Na 2 S·9H 2 O solution, 1.0mL Na 2 SeO 3 /Na 2 WO 4
solution, 1.0mL of sterile sodium dithionite solution, and 1.0mL of
sterile trace elements solution SL-10. Mix thoroughly. Aseptically and
anaerobically distribute into sterile tubes or flasks.
Use:  For the cultivation of Desulfotomaculum species.