Desulforhabdus amnigenus Medium

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulforhabdus amnigenus Medium

培养基配方
Composition per 1001.0mL:
Na2SO4 .......................................................................2.8g
Na2HPO4 ·2H2O................................................................0.53g
KH2PO4 .......................................................................0.41g
NaCl...........................................................................0.3g
NH4Cl .........................................................................0.3g
CaCl2 ·2H2O..................................................................0.11g
MgCl2 ·6H2O....................................................................0.1g
Resazurin .....................................................................0.5mg
Sodium propionate solution....................................................20.0mL
NaHCO3  solution .............................................................10.0mL
Wolfe’s vitamin solution.....................................................10.0mL
Na2S·9H2O solution...........................................................10.0mL
Selenite-tungstate solution....................................................1.0mL
Trace elements solution SL-10 .................................................1.0mL
Seven vitamin solution.........................................................1.0mL
pH 7.2–7.6 at 25°C
Sodium Propionate Solution:
Composition per 20.0mL:
Sodium propionate...............................................................2.0g
Preparation of Sodium Propionate Solution:  Add sodium pro-
pionate to distilled/deionized water and bring volume to 20.0mL. Mix
thoroughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 .........................................................................4.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O......................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl...............................................................10.0mg
p-Aminobenzoic acid............................................................5.0mg
Lipoic acid....................................................................5.0mg
Nicotinic acid.................................................................5.0mg
Riboflavin.....................................................................5.0mg
Thiamine·HCl..................................................................5.0mg
Calcium  DL -pantothenate......................................................5.0mg
Biotin.........................................................................2.0mg
Folic acid ....................................................................2.0mg
Vitamin B 12 ..................................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Selenite-Tungstate Solution:
Composition per liter:
NaOH............................................................................0.5g
Na2WO4 ·2H2O..................................................................4.0mg
Na2SeO3 ·5H2O.................................................................3.0mg
Preparation of 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.
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 Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
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 Seven Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas mixture. Add components, except sodium propi-
onate solution, NaHCO3  solution, selenite-tungstate solution, and
Na2S·9H2O solution, to distilled/deionized water and bring volume to
960.0mL. Mix thoroughly. Adjust pH to 7.2–7.6. Sparge with 80% N2
+ 20% CO2 gas mixture. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 20.0mL of sterile sodium
propionate solution, 10.0mL of sterile NaHCO3  solution, 1.0mL of
sterile selenite-tungstate solution, and 10.0mL of sterile Na2S·9H2O
solution. Mix thoroughly. Aseptically and anaerobically distribute into
sterile tubes or bottles.
Use: For the cultivation of Desulforhabdus amnigenus