Desulfacinum hydrothermale Medium (DSMZ Medium 875)

[所属分类:培养基配方] [发布时间:2020-6-2] [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

Desulfacinum hydrothermale Medium
(DSMZ Medium 875)

培养基配方
Composition per 1004mL:
Solution A...............................................................920.0mL
Soluiton C (NaHCO3 solution)..............................................50.0mL
Solution F................................................................13.0mL
Solution D (Seven vitamin solution) ......................................10.0mL
Solution E ...............................................................10.0mL
Soluiton B (Trace elements solution SL-10) ................................1.0mL
pH 7.0–7.3 at 25°C
Solution A:
Composition per 920mL:
NaCl.......................................................................10.4g
MgSO4 ·7H2O...............................................................2.72g
MgCl2 ·6H2O...............................................................2.24g
CaCl2 ·2H2O...............................................................0.56g
KCl........................................................................0.29g
NH4Cl .....................................................................0.1g
KH2PO4 ...................................................................0.08g
Resazurin ................................................................0.5mg
Preparation of Solution A:  Add components to distilled/deionized
water and bring volume to 920.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2 ·4H2O................................................................1.5g
CoCl2 ·6H2O ............................................................190.0mg
MnCl2 ·4H2O.............................................................100.0mg
ZnCl2 ....................................................................70.0mg
Na2MoO4 ·2H2O ...........................................................36.0mg
NiCl2 ·6H2O..............................................................24.0mg
H3BO3 .....................................................................6.0mg
CuCl2 ·2H2O ..............................................................2.0mg
HCl (25% solution)........................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add FeCl2 ·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min
at 15 psi pressure–121°C.
Solution C (NaHCO3 Solution:)
Composition per 100.0mL:
NaHCO3 .....................................................................5.0g
Preparation of Solution C (NaHCO3 Solution):  Add NaHCO3
to distilled/deionized water and bring volume to 100.0mL. Mix thor-
oughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
Solution D (Seven Vitamin Solution):
Composition per liter:
Pyridoxine hydrochloride ................................................300.0mg
Thiamine-HCl·2H2O.......................................................200.0mg
Nicotinic acid...........................................................200.0mg
Vitamin B 12 ............................................................100.0mg
Calcium pantothenate ....................................................100.0mg
p-Aminobenzoic acid.......................................................80.0mg
D (+)-Biotin..............................................................20.0mg
Preparation of Solution D (Seven Vitamin Solution): Add
components to distilled/deionized water and bring volume to 1.0L.
Sparge with 100% N2 . Mix thoroughly. Filter sterilize.
Solution E:
Composition per 10.0mL:
Na-lactate.................................................................2.5g
Preparation of Solution E:  Add Na-lactate to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution F:
Composition per 100.0mL:
Na2S·9H2O.................................................................3.0g
Preparation of Solution F:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add 50.0mL sterile solution C,
13.0mL sterile solution F, 10.0mL sterile solution D, 10.0mL sterile so-
lution E, and 1.0mL sterile solution B to 920.0mL sterile solution A.
Mix thoroughly. The pH of the completed medium should be 7.0–7.3.
Aseptically and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Desulfacinum hydrothermale.