MPOB Medium

[所属分类:培养基配方] [发布时间:2021-6-8] [发布人:网站管理员2] [阅读次数:] [返回]

MPOB Medium

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

Composition per 1012.0mL:

Disodium fumarate .......................................................................3.2g
Na 2 HPO 4 ·2H 2 O...........................................................................0.53g
KH 2 PO 4 .......................................................................................0.41g
NaCl..............................................................................................0.3g
NH 4 Cl ...........................................................................................0.3g
Yeast extract..................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................ 0.11g
MgCl 2 ·6H 2 O ...............................................................................0.10g
Resazurin...................................................................................0.5mg
NaHCO 3  solution.....................................................................20.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Selenite-tungstate solution.........................................................1.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0–7.2 at 25°C
NaHCO 3 Solution:
Composition per 20.0mL:
NaHCO 3 ........................................................................................4.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deion-
ized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 80%
N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to distilled/
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.

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.
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 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–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: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Prepare medium under 80% N 2 + 20%
CO 2 . Add components, except NaHCO 3  solution and Na 2 S·9H 2 O solu-
tion, to distilled/deionized water and bring volume to 970.0mL. Mix thor-
oughly. 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 sterile
NaHCO 3  solution and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thor-
oughly. Aseptically and anaerobically distribute into sterile tubes or flasks.
After inoculation, bring culture bottles to 0.7 bar 80% N 2 + 20% CO 2 over-
pressure.
Use:  For the cultivation of Syntrophobacter species.