MJ Medium (DSMZ Medium 1011)

[所属分类:培养基配方] [发布时间:2021-6-3] [发布人:网站管理员2] [阅读次数:] [返回]

MJ Medium
(DSMZ Medium 1011)

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

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
KH 2 PO 4 .......................................................................................0.14g
CaCl 2 ·2H 2 O...............................................................................0.014g
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 ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Thiosulfate solution .................................................................10.0mL
Bicarbonate solution................................................................10.0mL
pH 6.7 ± 0.2 at 25°C
Bicarbonate Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.5g
Preparation of Bicarbonate Solution:  Add components to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Thiosulfate Solution:
Composition per 10.0mL:
NaS 2 O 3 ·5H 2 O................................................................................1.5g
Preparation of Thiosulfate Solution:  Add components to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature.
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.
Preparation of Medium: Add components, except thiosulfate, bi-
carbonate, and vitamin solutions, to distilled/deionized water and bring
volume to 970.0mL. Mix thoroughly. Gently heat and bring to boiling.
Boil for 1 min. Cool to room temperature while sparging with a gas mix-
ture of 80% N 2 + 20% CO 2 . Dispense under the same atmosphere into
culture vessels. Fill up to a volume of 20% volume of vessel. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature under
an atmosphere of 0% N 2 + 20% CO 2 . Aseptically add sterile thiosulfate,
bicarbonate, and vitamin solutions. Mix thoroughly. Adjust pH to 6.7.
Aseptically dispense into tubes, flasks, or bottles. After inoculation pres-
surize vessels to 0.5 bar overpressure with 80% N 2 + 20% CO 2 gas mix-
ture. Add sterile air in an amount that is equivalent to a volume of 20%
of the headspace. After inoculation reduce medium with 10–20mg so-
dium dithionite per liter medium, added from a 5% solution freshly
prepared under N 2  and filter sterilized. Pressurize vessels to 2 bar over-
pressure with 80% H 2 and 20% CO 2 .
Use:  For the cultivation of Sulfurimonas autotrophica, Thiomicrospira
thermophila, and Desulfothermus okinawensis.