Desulfovibrio SHV Medium

[所属分类:培养基配方] [发布时间:2020-6-10] [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

Desulfovibrio SHV Medium

培养基配方
Composition per 1003.0mL:
Solution A.................................................................870.0mL
Solution C.................................................................100.0mL
Solution D..................................................................10.0mL
Solution E (Vitamin solution)...............................................10.0mL
Solution F..................................................................10.0mL
Solution B (Trace elements solution SL-10) ..................................1.0mL
Solution G (Selenite-tungstate solution).....................................1.0mL
Solution H (Seven vitamin solution) .........................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
NaCl...........................................................................7.0g
Na2SO4 ........................................................................3.0g
MgCl2 ·6H2O ..................................................................1.3g
KCl............................................................................0.5g
NH4Cl .........................................................................0.3g
KH2PO4 ........................................................................0.2g
CaCl2 ·2H2O .................................................................0.15g
Resazurin.....................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3–4 min. Allow to cool to room
temperature while gassing under 80% N2 + 20% CO2 . Continue gas-
sing until pH reaches below 6.0. Seal the flask under 80% N2 + 20%
CO2 . 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:
Composition per 100.0mL:
NaHCO3 ......................................................................5.0g
Preparation of Solution C:  Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Sparge with 80% N2 + 20% CO2 .
Solution D:
Composition per 10.0mL:
Sodium lactate ...............................................................4.0g
Preparation of Solution D:  Add sodium lactate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E (Vitamin Solution):
Composition per liter:
Pyridoxine·HCl..............................................................10.0mg
Calcium  DL -pantothenate.....................................................5.0mg
Lipoic acid ..................................................................5.0mg
Nicotinic acid................................................................5.0mg
p-Aminobenzoic acid...........................................................5.0mg
Riboflavin ...................................................................5.0mg
Thiamine·HCl ................................................................5.0mg
Biotin .......................................................................2.0mg
Folic acid....................................................................2.0mg
Vitamin B 12 .................................................................0.1mg
Preparation of Solution E (Vitamin Solution): Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O.....................................................................0.4g
Preparation of Solution F:  Add Na2S·9H2O 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 G (Selenite-Tungstate Solution):
Composition per liter:
NaOH............................................................................0.5g
Na2WO4 ·2H2O..................................................................4.0mg
Na2SeO3 ·5H2O.................................................................3.0mg
Preparation of Solution G (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution H (Seven Vitamin Solution) :
Composition per liter:
Pyridoxine·HCl................................................................0.3g
Thiamine·HCl .................................................................0.2g
Nicotinic acid.................................................................0.2g
Calcium  DL -pantothenate......................................................0.1g
Vitamin B 12 ..................................................................0.1g
p-Aminobenzoic acid..........................................................80.0mg
Biotin ......................................................................20.0mg
Preparation of Solution H (Seven Vitamin Solution):  Add
components to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly.
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, solution
F, solution G, and solution H, in that order. Mix thoroughly. Anaerobi-
cally distribute into sterile tubes or flasks under 80% N2 + 20% CO2 .
Use: For the cultivation of Desulfovibrio species.