Pelotomaculum Medium (DSMZ Medium 960)

[所属分类:培养基配方] [发布时间:2021-7-1] [发布人:网站管理员2] [阅读次数:] [返回]

Pelotomaculum Medium
(DSMZ Medium 960)

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

Composition per liter:

NaHCO 3 ........................................................................................2.5g
NH 4 Cl .........................................................................................0.54g
MgCl 2 ·6H 2 O..................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
KH 2 PO 4 .......................................................................................0.14g
Yeast extract..................................................................................0.1g
Resazurin ...................................................................................0.5mg
Na-pyruvate solution................................................................20.0mL
Cysteine solution......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Vitamin solution.........................................................................5.0mL
Trace elements solution.............................................................1.0mL
Selenite-tungstate solution.........................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Na-Pyruvate Solution :
Composition per 20.0mL:
Na-pyruvate ..................................................................................2.2g
Preparation of Na-Pyruvate  Solution : Add Na-pyruvate to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
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.
Cool to room temperature.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.3g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H 2 O 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. Cool to room temperature.
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.
Trace Elements Solution:
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 : 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 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Add components, except Na-pyruvate so-
lution, cysteine solution, and Na 2 S·9H 2 O solution, to distilled/deion-
ized water and bring volume to 960.0mL. Mix thoroughly. Sparge with
80% N 2 + 20% CO 2 . Equilibrate with this gas mixture to reach pH 7.0.
Distribute into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add per liter of medium
20.0mL sterile Na-pyruvate solution, 10.0mL sterile cysteine solution,
and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly.
Use:  For the cultivation of Pelotomaculum thermopropionicum.