Desulfovibrio halophilus Medium

[所属分类:培养基配方] [发布时间:2020-6-9] [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 halophilus Medium

培养基配方
Composition per 1154.0mL:
Solution A...................................................................1.0L
Solution H.................................................................67.0mL
Solution D.................................................................50.0mL
Soultion I.................................................................13.0mL
Solution E ................................................................10.0mL
Solution G.................................................................10.0mL
Solution C (Selenite-tungstate solution)....................................2.0mL
Solution B (Trace elements solution SL-10) .................................1.0mL
Solution F..................................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Solution A:
Composition per liter:
Na2SO4 ......................................................................4.0g
NH4Cl ......................................................................0.25g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O.................................................................0.1g
Preparation of Solution A:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Continue boiling for 3–4 min. Allow to cool to room tem-
perature while gassing under 80% N2 + 20% CO2 . Continue gassing
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. Gas under 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution C (Selenite-Tungstate Solution):
Composition per liter:
NaOH.......................................................................0.5g
Na2WO4 ·2H2O.............................................................4.0mg
Na2SeO3 ·5H2O............................................................3.0mg
Preparation of Solution C (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Gas under 100% N2 . Autoclave for 15 min at 15
psi pressure–121°C.
Solution D:
Composition per 50.0mL:
NaHCO3 ..................................................................2.5g
Preparation of Solution D:  Add NaHCO3 to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Gas under 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E:
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:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N2 .
Filter sterilize.
Solution F:
Composition per 10.0mL:
Vitamin B 12 .........................................................0.5mg
Preparation of Solution F:  Add vitamin B 12 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Filter sterilize.
Solution G:
Composition per 80.0mL:
Sodium-( L )-lactate..................................................2.25g
Preparation of Solution G : Add sodium-( L )-lactate to distilled/de-
ionized water and bring volume to 80.0mL. Mix thoroughly. Gas under
100% N2 . In a closed bottle, heat in a boiling water bath. Shake until
stearic acid dissolves. Autoclave for 15 min at 15 psi pressure–121°C.
On storage, solution will solidify and should be remelted before use.
Solution H:
Composition per liter:
NaCl....................................................................70.4g
MgCl2 ·6H2O ...........................................................3.0g
CaCl2 ·2H2O ...........................................................2.2g
Preparation of Solution H: Add components to distilled/deionized
water and bring volume to 80.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution I:
Composition per 20.0mL:
Na2S·9H2O..............................................................0.15g
Preparation of Solution I:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: To 1.0L of sterile solution A, add in the fol-
lowing order: 1.0mL of sterile solution B, 2.0mL of sterile solution C,
50.0mL of sterile solution D, 10.0mL of sterile solution E, 1.0mL of ster-
ile solution F, 10.0mL of sterile solution G, 67.0mL of sterile solution H,
and 13.0mL of sterile solution I. Mix thoroughly. Final pH of medium
should be 7.2. Prior to inoculation, add 10.0–20.0mg of sodium dithion-
ate to 1.0L of medium.
Use:  For the cultivation and maintenance of Desulfovibrio halophilus.