Desulfotomaculum acetoxidans 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

Desulfotomaculum acetoxidans Medium

培养基配方
Composition per 1020.0mL:
Solution A....................................................................1.0L
Na2S·9H2O solution.........................................................10.0mL
Wolfe’s vitamin solution...................................................10.0mL
Solution A:
Composition per liter:
NaHCO3 .......................................................................4.5g
Na2SO4 ......................................................................2.84g
Sodium acetate................................................................1.4g
Sodium butyrate...............................................................1.4g
NaCl.........................................................................1.17g
Yeast extract.................................................................1.0g
MgCl2 ·6H2O .................................................................0.4g
KCl...........................................................................0.3g
NH4Cl .......................................................................0.27g
KH2PO4 .......................................................................0.2g
CaCl2 ·2H2O .................................................................0.15g
Resazurin ....................................................................0.5mg
Trace elements solution SL-7..................................................1.0mL
Trace Elements Solution SL-7:
Composition per 1010.0mL:
FeCl2 ·4H2O...................................................................1.5g
CoCl2 ·6H2O................................................................190.0mg
MnCl2 ·4H2O ...............................................................100.0mg
ZnCl2 .......................................................................70.0mg
H3BO3 .......................................................................62.0mg
Na2MoO4 ·2H2O...............................................................36.0mg
NiCl2 ·6H2O.................................................................24.0mg
CuCl2 ·2H2O.................................................................17.0mg
Hydrochloric acid, 25%.......................................................10.0mL
Preparation of Trace Elements Solution SL-7: 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.
Preparation of Solution A: Prepare and dispense medium under
80% N2  + 20% CO2 . Add components, except NaHCO3 and bring vol-
ume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool to
room temperature while sparging with 80% N2 + 20% CO2 . Add
NaHCO3 . Mix thoroughly. Continue sparging with 80% N2 + 20% CO2
until pH stabilizes at 6.9–7.1. Anaerobically distribute into tubes in
10.0mL amounts under an atmosphere of 80% N2 + 20% CO2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O..................................................................0.36g
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.
Filter sterilize. Sparge with 100% N2 .
Preparation of Medium: Aseptically and anaerobically add 0.1mL
of sterile Na2S·9H2O solution and 0.1mL of sterile Wolfe’s vitamin so-
lution to each tube containing 10.0mL of solution A. Prepare immedi-
ately prior to use.
Use: For the cultivation of Desulfotomaculum acetoxidans.