MV Medium

[所属分类:培养基配方] [发布时间:2021-6-11] [发布人:网站管理员2] [阅读次数:] [返回]

MV Medium

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

Composition per 1001.0mL:

Na 2 SO 4 ..........................................................................................2.0g
MgSO 4 ·7H 2 O................................................................................1.0g
Na 2 S 2 O 3 5H 2 O...............................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
Yeast extract..................................................................................1.0g
KH 2 PO 4 .........................................................................................0.5g
CaCl 2 ·2H 2 O ..................................................................................0.1g
Resazurin ...................................................................................0.5mg
Wolfe’s vitamin solution..........................................................10.0mL
Sodium malate solution ...........................................................10.0mL
Sodium pyruvate solution........................................................10.0mL
NaHCO 3  solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0–7.2 at 25°C
Sodium Malate Solution:
Composition per 100.0mL:
Sodium malate..............................................................................1.0g
Preparation of Sodium Malate Solution:  Add sodium malate to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Sodium Pyruvate Solution:
Composition per 100.0mL:
Sodium pyruvate...........................................................................1.0g
Preparation of Sodium Pyruvate Solution:  Add sodium pyru-
vate 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.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. 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................................................................................0.75μg
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 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.
Sparge with 100% N 2 . Filter sterilize.
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 and dispense medium under
100% N 2 . Add components, except Wolfe’s vitamin solution, sodium
malate solution, sodium pyruvate solution, NaHCO 3  solution,
Na 2 S·9H 2 O solution, and trace elements solution SL-10, to distilled/
deionized water and bring volume to 950.0mL. Mix thoroughly. Adjust
pH to 7.0–7.2. Gently heat and bring to boiling. Cool while sparging
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Asep-
tically and anaerobically add 10.0mL of sterile Wolfe’s vitamin solu-
tion, 10.0mL of sterile sodium malate solution, 10.0mL of sterile
sodium pyruvate solution, 10.0mL of sterile NaHCO 3  solution, 10.0mL
of sterile Na 2 S·9H 2 O solution, and 1.0mL of sterile trace elements so-
lution SL-10. Mix thoroughly. Aseptically and anaerobically distribute
into sterile tubes or bottles.
Use: For the cultivation of Desulfotomaculum orientis.