16AV Medium (DSMZ Medium 298f)

[所属分类:培养基配方] [发布时间:2019-11-15] [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

16AV Medium
(DSMZ Medium 298f)
Composition per liter:
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl2 ·6H2O..................................................................................0.4g
NH4Cl .........................................................................................0.25g
KH2PO4 .........................................................................................0.2g
CaCl2 ·2H2O.................................................................................0.15g
Resazurin ...................................................................................1.0mg
NaHCO3  solution .....................................................................10.0mL
Butanediol solution..................................................................10.0mL
Na2S·9H2O solution.................................................................10.0mL
Yeast extract solution...............................................................10.0mL
Galactose solution....................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na2S·9H 2 O Solution :
Composition per 10.0mL:
Na2S·9H2O..................................................................................0.36g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Filter sterilize.
Butanediol Solution:
Composition per 10.0mL:
2,3 butanediol................................................................................0.9g
Preparation of Butanediol Solution: Add butanediol to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Galactose Solution:
Composition per 10.0mL:
Galactose.......................................................................................2.0g
Preparation of Galactose Solution: Add galactose to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
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 FeCl 2 ·4H 2 O
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% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................1.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solu-
tion, butanediol solution, Na 2 S·9H 2 O solution, galactose solution, yeast
extract solution, and trace elements solution SL-10, to distilled/deion-
ized water and bring volume to 949.0mL. Mix thoroughly. Adjust pH to
7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically add 10.0mL NaHCO 3
solution, 10.0mL butanediol solution, 10.0mL Na 2 S·9H 2 O solution,
10.0mL galactose soltuion, 10.0mL yeast extract solution, and 1.0mL
trace elements solution SL-10. Mix thoroughly. Aseptically and anaero-
bically distribute into sterile tubes or bottles. After inoculation, flush and
repressurize the gas head space of culture bottles with sterile 80% N 2 +
20% CO 2 to 1 bar overpressure.
Use:  For the cultivation of unclassified bacterium DSM 8385.