Nitratiruptor and Nitratifactor Medium (DSMZ Medium 1024)

[所属分类:培养基配方] [发布时间:2021-6-18] [发布人:网站管理员2] [阅读次数:] [返回]

Nitratiruptor and Nitratifactor Medium
(DSMZ Medium 1024)

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, elemental...........................................................................3.0g
Na 2 S 2 O 3 ·5H 2 O..............................................................................1.0g
NaNO 3 ..........................................................................................1.0g
Bicarbonate solution................................................................10.0mL
Vitamin solution.......................................................................10.0mL
DMJ synthetic seawater ...............................................................1.0L
pH 7.0 ± 0.2 at 25°C
Bicarbonate Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.0g
Preparation of Bicarbonate Solution:  Add NaHCO 3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
DMJ Synthetic Seawater:
Composition per liter:
NaCl ...........................................................................................30.0g
MgCl 2 ·6H 2 O................................................................................4.18g
MgSO 4 · 7H 2 O................................................................................3.4g
KCl..............................................................................................0.33g
NH 4 Cl ........................................................................................0.25g
K 2 HPO 4 ......................................................................................0.14g
CaCl 2 ·2H 2 O.................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O ............................................................... 0.01g
NiCl 2 ·6H 2 O................................................................................0.5mg
Na 2 SeO 3 ·5H 2 O...........................................................................0.5mg
Trace elements solution SL-10 ................................................10.0mL
Trace Elements Solution SL-10:
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 SL-10:  Add nitrilotri-
acetic acid to 500.0mL of distilled/deionized water. Dissolve by adjust-
ing pH to 6.5 with KOH. Add remaining components. Add distilled/
deionized water to 1.0L. Mix thoroughly. Adjust pH to 7.0.
Preparation of DMJ Synthetic Seawater:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C . Cool to room temper-
ature.
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.
Preparation of Medium: Add components, except sulfur, bicar-
bonate solution, and vitamin solution to seawater and bring volume to
1.0L. Dispense into serum bottles. Autoclave for 15 min at 15 psi pres-
sure–121°C under an atmosphere of air. Sterilize sulfur separately in
screw-capped tubes by steaming in a water bath for 3 hr on each of 3
successive days. Aseptically add the sterilized sulfur, bicarbonate, and
vitamin solutions. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 .
Seal the serum tubes with butyl rubber stoppers. Increase the 80% H 2
+ 20% CO 2 gas phase pressure to 300 kPa.
Use: For the cultivation of Nitratiruptor spp. and Nitratifactor spp.