Rhodocyclus purpureus Medium (DSMZ Medium 44)

[所属分类:培养基配方] [发布时间:2021-8-4] [发布人:网站管理员2] [阅读次数:] [返回]

Rhodocyclus purpureus Medium
(DSMZ Medium 44)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1056.9 mL:

Na 2 -succinate................................................................................1.0g
(NH 4 )-acetate................................................................................0.5g
KH 2 PO 4 .........................................................................................0.5g
MgSO 4 ·7H 2 O................................................................................0.4g
NaCl..............................................................................................0.4g
NH 4 Cl ...........................................................................................0.4g
Yeast extract..................................................................................0.3g
CaCl 2 ·2H 2 O ................................................................................0.05g
Ferric citrate solution.................................................................5.0mL
Trace elements solution SL-6....................................................1.0mL
Ethanol.......................................................................................0.5mL
Vitamin B 12  solution ..................................................................0.4mL
Neutralized sulfide solution.................................................... variable
pH 6.8 ± 0.2 at 25°C
Trace Elements Solution SL-6:
Composition per liter:
MnCl 2 ·4H 2 O .................................................................................0.5g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O..................................................................................0.2g
ZnSO 4 ·7H 2 O................................................................................ 0.1g
Na 2 MoO 4 ·2H 2 O......................................................................... 0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2 ·2H 2 O................................................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Vitamin B 12 Solution:
Composition per 100.0mL:
Vitamin B 12 ..............................................................................10.0mg
Preparation of Vitamin B 12 Solution:  Add vitamin B 12 to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Sparge under 100% N 2  gas for 3 min.
Ferric Citrate Solution:
Composition per 10.0mL:
Ferric citrate.............................................................................10.0mg
Preparation of Ferric Citrate Solution : Add ferric citrate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge under 100% N 2 gas for 3 min.
Neutralized Sulfide Solution:
Composition per 100.0mL:
Na 2 S·9H 2 O....................................................................................1.5g
Preparation of Neutralized Sulfide Solution : Add Na 2 S·9H 2 O
to distilled/deionized water in a 250.0mL screw-capped bottle fitted
with a butyl rubber septum and bring volume to 100.0mL. Add a mag-
netic stir bar. Mix thoroughly. Sparge under 100% N 2 gas for 3 min.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature. Adjust pH to about 7.0 with sterile 2M H 2 SO 4 . Do not open the
bottle to add H 2 SO 4 ; use a sterile syringe. Stir the solution continuously
to avoid precipitation of elemental sulfur. The final solution should be
clear and yellow in color.
Preparation of Medium: Add components, except neutralized sul-
fide solution, to 1050.0mL distilled/deionized water. Mix thoroughly.
Gently heat and bring to boiling. Boil for 3–4 min under a stream of
100% N 2 . Distribute 45.0mL of the prepared medium into 50.0mL
screw-capped tubes that have been flushed with 100% N 2 . Autoclave
for 15 min at 15 psi pressure–121°C. Cool to room temperature. Before
inoculation, aseptically and anaerobically add 0.25–0.50mL of neutral-
ized sulfide solution. Sterile syringes are used to inoculate and remove
samples. Incubate in the light using a tungsten lamp.
Use: For the cultivation and maintenance of brown and other oxygen-
sensitive Rhodospirillaceae.