Desulfovibrio sulfodismutans 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 sulfodismutans Medium

培养基配方
Composition per 1002.0mL:
Solution A.................................................................920.0mL
Solution C..................................................................50.0mL
Solution D..................................................................10.0mL
Solution F..................................................................10.0mL
Solution G..................................................................10.0mL
Solution B (Trace elements solution SL-10) ..................................1.0mL
Solution E ..................................................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 920.0mL:
NaCl..........................................................................1.0g
KCl...........................................................................0.5g
MgCl2 ·6H2O .................................................................0.4g
NH4Cl ........................................................................0.3g
KH2PO4 .......................................................................0.2g
CaCl2 ·2H2O.................................................................0.15g
Preparation of Solution A: Prepare and dispense solution anaero-
bically under 80% N2 + 20% 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. Cap with rubber stoppers. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 45°–50°C.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2 ·4H2O..................................................................1.5g
CoCl2 ·6H2O ................................................................0.19g
MnCl2 ·4H2O..................................................................0.1g
ZnCl2 .......................................................................0.07g
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% N 2 for 20 min.
Solution C:
Composition per 50.0mL:
NaHCO3 .......................................................................2.5g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize.
Aseptically gas under 80% N2 + 20% CO2 for 20 min.
Solution D:
Composition per 10.0mL:
Na-acetate·3H2O...............................................................0.3g
Preparation of Solution D: Prepare and dispense solution anaero-
bically under 80% N2 + 20% CO2 . Add Na-acetate·3H2O to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Cap
with rubber stopper. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 45°–50°C.
Solution E:
Composition per liter:
Calcium pantothenate ........................................................0.05mg
Biotin ......................................................................0.01mg
Preparation of Solution E: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Asep-
tically gas under 100% N2 for 20 min.
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. 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 G:
Composition per 10.0mL:
Na2S2O5 .......................................................................1.05g
Preparation of Solution G: Add Na2S2O5 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Aseptically gas under 100% N2 for 20 min. Prepare solution freshly.
Preparation of Medium: To 920.0mL of cooled, sterile solution A,
aseptically and anaerobically add in the following order: 1.0mL of ster-
ile solution B, 50.0mL of sterile solution C, 10.0mL of sterile solution
D, 1.0mL of sterile solution E, 10.0mL of sterile solution F, and
10.0mL of sterile solution G. Mix thoroughly. Aseptically and anaero-
bically distribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Desulfovibrio sulfodismu-
tans.