Desulfovibrio sapovorans 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 sapovorans Medium

培养基配方
Composition per 1009.0mL:
Solution A..............................................................850.0mL
Solution C..............................................................100.0mL
Solution G...............................................................20.0mL
Solution D...............................................................10.0mL
Solution E (Wolfe’s vitamin solution)...................................10.0mL
Solution H...............................................................10.0mL
Solution F................................................................6.6mL
Solution B (Trace elements solution SL-10) ...............................1.0mL
Solution I ...............................................................0.4mL
pH 7.7 ± 0.2 at 25°C
Solution A:
Composition per 920.0mL:
Na2SO4 ....................................................................3.0g
NaCl.......................................................................1.0g
KCl........................................................................0.5g
MgCl2 ·6H2O ..............................................................0.4g
NH4Cl .....................................................................0.3g
KH2PO4 ....................................................................0.2g
CaCl2 ·2H2O .............................................................0.15g
Resazurin.................................................................0.5mg
Preparation of Solution A: Prepare and dispense solution anaero-
bically under 90% N2 + 10% CO2 . Add components to distilled/deion-
ized water and bring volume to 920.0mL. Mix thoroughly. Gently heat
and bring to boiling. Continue boiling until resazurin turns colorless,
indicating reduction, and a pH of 6.0 is reached. Cap with rubber stop-
pers. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2 ·4H2O.................................................................1.5g
CoCl2 ·6H2O................................................................0.19g
MnCl2 ·4H2O ...............................................................0.10g
ZnCl2 .....................................................................0.070g
Na2MoO4 ·2H2O.............................................................0.036g
NiCl2 ·6H2O...............................................................0.024g
H3BO3 ......................................................................6.0mg
CuCl2 ·2H2O................................................................2.0mg
HCl (25% solution).........................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add the FeCl2 ·4H2O to 10.0mL of HCl solution. Mix thoroughly.
Bring volume to approximately 900.0mL with distilled/deionized wa-
ter. Mix thoroughly. Adjust pH to 6.0 with NaOH. Bring volume to
1.0L with distilled/deionized water. Filter sterilize. Aseptically gas un-
der 100% N2 for 20 min.
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.
Aseptically gas under 90% N2 + 10% CO2 for 20 min.
Solution D:
Composition per 10.0mL:
Sodium butyrate..............................................................0.7g
Sodium caproate .............................................................0.3g
Sodium octanoate............................................................0.15g
Preparation of Solution D: Prepare and dispense solution anaero-
bically under 90% N2 + 10% CO2 . Add components to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Cap with a
rubber stopper. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C.
Solution E (Wolfe’s Vitamin Solution):
Composition per liter:
Pyridoxine·HCl............................................................0.01g
Thiamine·HCl .............................................................5.0mg
Riboflavin ................................................................5.0mg
Nicotinic acid.............................................................5.0mg
Calcium pantothenate ......................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Thioctic acid .............................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Cyanocobalamin ............................................................0.1mg
Preparation of Solution E (Wolfe’s Vitamin Solution): Add
components to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Filter sterilize. Aseptically gas under 100% N2 for 20 min.
Solution F:
Composition per 6.6mL:
AlCl3 ·6H2O (4.9% solution)...............................................5.0mL
Na2CO3 (10.6% solution)....................................................1.6mL
Preparation of Solution F: Combine both solutions. Mix thor-
oughly. Gas with 100% N2 . Cap with a rubber stopper. Autoclave for
15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution G:
Composition per 10.0mL:
Rumen fluid, clarified.....................................................20.0mL
Preparation of Solution G: Gas rumen fluid under 100% N2 for 20
min. Cap with a rubber stopper. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Solution H:
Composition per 10.0mL:
Na2S·9H2O...................................................................0.4g
Preparation of Solution H: Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Gas under 100% N2 for 20 min.
Cap with a rubber stopper. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Solution I:
Composition per 10.0mL:
Na2S2O4 ......................................................................0.5g
Preparation of Solution I: Add Na2S2O4 to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Asep-
tically gas under 100% N2 for 20 min. Prepare solution freshly.
Preparation of Medium: Prepare and dispense medium under 90%
N2 + 10% CO2 . To 850.0mL of cooled, sterile solution A, aseptically and
anaerobically add in the following order: 1.0mL of sterile solution B,
100.0mL of sterile solution C, 10.0mL of sterile solution D, 10.0mL of
sterile solution E, 6.6mL of sterile solution F, 20.0mL of sterile solution G,
and 10.0mL of sterile solution H. Mix thoroughly. Immediately prior to in-
oculation, aseptically and anaerobically add 0.4mL of sterile solution I.
Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes
or flasks.
Use:  For the cultivation and maintenance of Desulfovibrio sapovorans.