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山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Acid Rhodospirillaceae Medium
Composition per 1050.0 mL:
Ammonium acetate.......................................................................1.5g
KH 2 PO 4 .........................................................................................0.5g
MgSO 4 .7H 2 O............................................................................... 0.4g
NaCl..............................................................................................0.4g
NH 4 Cl ...........................................................................................0.4g
Disodium succinate.....................................................................0.25g
Yeast extract..................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.05g
Ferric citrate solution.................................................................5.0mL
Trace elements solution SL-6 ....................................................1.0mL
Vitamin B 12 solution..................................................................0.4mL
Neutralized sulfide solution....................................................variable
pH 5.7 ± 0.2 at 25°C
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. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
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. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
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 7.3 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 distilled/deionized water and bring volume to
1050.0mL. 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 neutralized sulfide solution.
Use: For the cultivation and maintenance of members of the family
Rhodospirillaceae, including Rhodomicrobium vannielii and Rho-
dopseudomonas acidophila.