AMH (DSMZ Medium 1110)

[所属分类:培养基配方] [发布时间:2019-10-8] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

AMH
(DSMZ Medium 1110)
Composition per liter:
NaCl ...........................................................................................20.0g
Sulfur, powdered ..........................................................................5.0g
MgCl 2 ·6H 2 O..................................................................................3.0g
KCl................................................................................................0.5g
NH 4 Cl ........................................................................................ 0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Resazurin ..................................................................................0.5mg
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
NaHCO 3  solution .....................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
Selenite-tungstate solution ........................................................1.0mL
pH 7.0 ± 0.2 at 25°C
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3  to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% H 2 . Autoclave 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....................................................................................0.5g
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.
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.
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 . Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except bicarbonate so-
lution, sulfide solution, sulfur, and vitamin solution, to distilled/deion-
ized water and bring volume to 970.0mL. Mix thoroughly. Gently heat
and bring to boiling. Boil for several minutes. Cool to room tempera-
ture while sparging with 80% H 2 + 20% CO 2 . Add the bicarbonate so-
lution, sulfide solution, and vitamin solution. Distribute into scew-
capped tubes or bottles. Autoclave for 15 min at 15 psi pressure–
121°C. Place the sulfur in screw-capped tubes or bottles. Autoclave for
15 min at 8 psi pressure–112°C. Before use, aseptically and anoxically
layer the sulfur onto the surface of sterile liquid basal medium. Adjust
the final pH to 7.0. After inoculation, pressurize culture vials to 1 bar
overpressure with 80% H 2 and 20% CO 2 gas mixture.
Use: For the maintenance or cultivation of Nautilia profundicola.