DSIC Medium, Modified (DSMZ Medium 747)

[所属分类:培养基配方] [发布时间:2020-6-28] [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

DSIC Medium, Modified
(DSMZ Medium 747)

培养基配方
Composition per 994.0mL:
Solution A............................................................910.0mL
Solution B.............................................................70.0mL
Solution C.............................................................14.0mL
pH 7.0–7.1 at 25°C
Solution A:
Composition per 960.0mL:
NaCl...................................................................125.0g
K2SO4 ...................................................................2.5g
Na-acetate...............................................................2.0g
Yeast extract...........................................................0.75g
KH2PO4 ..................................................................0.6g
NH4Cl ...................................................................0.5g
Na2S2O3 ·5H2O ..........................................................0.1g
MOPS buffer............................................................10.0mL
Vitamin B 12 solution...................................................1.0mL
Trace elements solution SL-10...........................................1.0mL
Vitamin B 12 Solution:
Composition per 100.0mL:
Vitamin B 12 ..........................................................10.0mg
Vitamin B 12 Solution:  Add vitamin B 12 to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Sparge under 100% N2
gas for 3 min.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O............................................................1.5g
CoCl2 ·6H2O.........................................................190.0mg
MnCl2 ·4H2O ........................................................100.0mg
ZnCl2 ................................................................70.0mg
Na2MoO4 ·2H2O........................................................36.0mg
NiCl2 ·6H2O..........................................................24.0mg
H3BO3 .................................................................6.0mg
CuCl2 ·2H2O...........................................................2.0mg
HCl (25% solution)....................................................10.0mL
Preparation of Trace Elements Solution SL-10: 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. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
MOPS Buffer:
Composition per 10.0mL:
MOPS [3-(N-morpholino) propane
sulfonic acid]...........................................................2.1g
Na-acetate ..............................................................0.3g
EDTA.....................................................................0.1g
Preparation of MOPS Buffer: Add components to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Adjust to pH 7.2. Filter sterilize.
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 960.0mL. Mix thoroughly. Sparge with
100% N2 . Gently heat and bring to boiling while continuing to sparge
with 100% N2 . Distribute about 13mL aliquots in 15mL Hungate tubes.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution B:
Composition per 70.0mL:
MgCl2 ·6H2O............................................................10.0g
CaCl2 ·2H2O.............................................................0.2g
Preparation of Solution B: Add components to distilled/deionized
water and bring volume to 70.0mL. Mix thoroughly. Adjust pH to 7.0.
Sparge with 100% N2 . Gently heat and bring to boiling while continuing
to sparge with 100% N2 . Distribute into a screw-capped bottle. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution C:
Composition per 14.0mL:
NaHCO3 ..................................................................1.0g
Preparation of Solution C:  Add NaHCO3 to distilled/deionized
water and bring volume to 14.0mL. Mix thoroughly. Sparge with 80%
N2 + 20% CO2 . Filter sterilize.
Preparation of Medium: Inject 1.0 ml of solution B and 0.2 ml of
solution C in each tube of solution A.
Use:  For the cultivation of Rhodovibrio sodomensis=Rhodospirillum
sodomense.