Nautilia Medium (DSMZ Medium 946)

[所属分类:培养基配方] [发布时间:2021-6-17] [发布人:网站管理员2] [阅读次数:] [返回]

Nautilia Medium
(DSMZ Medium 946)

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

Composition per liter:

Sulfur..........................................................................................10.0g
NH 4 Cl .........................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
Resazurin ...................................................................................0.5mg
Synthetic seawater, concentrated...........................................500.0mL
NaHCO 3 solution.....................................................................50.0mL
Na 2 S·9H 2 O solution.................................................................20.0mL
Sodium formate solution..........................................................15.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Selenite-tungstate solution.........................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Synthetic Seawater, Concentrated:
Composition per 500.0mL:
NaCl............................................................................................55.4g
MgSO 4 ·7H 2 O..............................................................................14.0g
MgCl 2 ·6H 2 O ............................................................................... 11.0g
CaCl 2 ·2H 2 O ..................................................................................1.5g
KCl................................................................................................1.3g
NaBr..............................................................................................0.2g
H 3 BO 3 .........................................................................................0.06g
SrCl 2 ·6H 2 O .................................................................................0.03g
Na 3 -citrate ...............................................................................20.0mg
KI.............................................................................................. 0.1mg
Preparation of Synthetic Seawater, Concentrated: Add com-
ponents to distilled/deionized water and bring volume to 500.0mL. Mix
thoroughly.
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.
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.
Sodium Formate Solution:
Composition per 50.0mL:
Na-formate..................................................................................10.0g
Preparation of Sodium Formate Solution: Add sodium formate
to distilled/deionized water and bring volume to 50.0mL. Mix thor-
oughly. Sparge with 100% N 2 . Filter sterilize.
Na 2 S·9H 2 O Solution :
Composition per 20.0mL:
Na 2 S·9H 2 O....................................................................................0.6g
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 20.0mL. 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 ........................................................................................5.0g
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.
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 Sulfur:  Sterilize sulfur by steaming for 3 h on each
of 3 successive days.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except sulfur,
NaHCO 3  solution, sodium formate solution, Na 2 S·9H 2 O solution, vita-
min solution, selenite-tungstate solution, and trace elements solution,
to distilled/deionized water and bring volume to 894.0mL. Mix thor-
oughly. Adjust pH to 6.8. Sparge with 80% N 2 + 20% CO 2 . Autoclave
for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add
10.0g steam sterilized sulfur, 50.0mL NaHCO 3  solution, 15.0mL sodi-
um formate solution, 20.0mL Na 2 S·9H 2 O solution, 10.0mL vitamin so-
lution, 1.0mL selenite-tungstate solution, and 10.0mL trace elements
solution. Mix thoroughly. Adjust pH to 6.8. Aseptically and anaerobical-
ly distribute into sterile tubes or bottles.
Use:  For the cultivation of Caldithrix abyssi and Nautilia lithotrophica.