Natroniella Medium (DSMZ Medium 784)

[所属分类:培养基配方] [发布时间:2021-6-16] [发布人:网站管理员2] [阅读次数:] [返回]

Natroniella Medium
(DSMZ Medium 784)

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

Composition per liter:

Na 2 CO 3  .......................................................................................68.3g
NaCl............................................................................................15.7g
NH 4 Cl ...........................................................................................1.0g
KCl............................................................................................... 0.2g
KH 2 PO 4 .........................................................................................0.2g
Yeast extract..................................................................................0.2g
MgCl 2 ·6H 2 O .................................................................................0.1g
Resazurin ...................................................................................0.5mg
NaHCO 3  solution ...................................................................100.0mL
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Ethanol solution.......................................................................10.0mL
Trace elements solution SL-11...................................................1.0mL
pH 9.7–10.0 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.0g
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.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO 3 ......................................................................................38.3g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Filter sterilize.
Ethanol Solution:
Composition per 10.0mL:
Ethanol.......................................................................................5.0mL
Preparation of Ethanol Solution:  Add ethanol to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 2 . Filter sterilize.
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 SL-4:
Composition per liter:
EDTA............................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.2g
Trace elements solution SL-6 ................................................100.0mL
Trace Elements Solution SL-6:
Composition per liter:
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O ..................................................................................0.2g
ZnSO 4 ·7H 2 O.................................................................................0.1g
MnCl 2 ·4H 2 O................................................................................0.03g
Na 2 MoO 4 ·H 2 O ............................................................................0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2· ·2H 2 O................................................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Ad-
just pH to 3.4.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except vitamin solution, NaHCO 3 solu-
tion, ethanol solution, and Na 2 S·9H 2 O solution, to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly. Gently heat and
bring to boiling. Cool to room temperature while sparging with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Aseptically and anaerobically add 10.0mL sterile vitamin solution,
100.0mL of sterile NaHCO 3 solution, 10.0mL sterile ethanol solution,
and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Adjust pH
to 9.7–10.0. Aseptically and anaerobically distribute into sterile tubes
or flasks.
Use:  For the cultivation of Natroniella acetigena (Acetohalobium sp.).