Acetivibrio Desulfovibrio Medium (LMG Medium 105)

[所属分类:培养基配方] [发布时间:2019-9-18] [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

Acetivibrio Desulfovibrio Medium
(LMG Medium 105)
Composition per liter:
Solution A..............................................................................869.0mL
Solution C..............................................................................100.0mL
Solution D................................................................................10.0mL
Solution E ................................................................................10.0mL
Solution F.................................................................................10.0mL
Solution B..................................................................................1.0mL
pH 7.7 ± 0.2 at 25°C
Solution A:
Composition per 869.0mL:
Na 2 SO 4 ..........................................................................................3.0g
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.4g
NH 4 Cl ...........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Preparation of  Solution A: Add components to distilled/deionized
water and bring volume to 869.0mL. Mix thoroughly. Prepare and au-
toclave part A under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C. Cool to room temperature.
Solution B:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ................................................................................0.19g
MnCl 2 ·4H 2 O..................................................................................0.1g
ZnCl 2 ...........................................................................................0.07g
H 3 BO 3 .........................................................................................0.06g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.04g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2 ·2H 2 O ................................................................................0.02g
HCl, 25% .................................................................................10.0mL
Preparation of  Solution B: Add the FeCl 2 ·4H 2 O to the HCl. Add
distilled/deionized water and bring volume to 1.0L. Add remaining
components. Mix thoroughly. Autoclave under 100% N 2 for 15 min at
15 psi pressure–121°C. Cool to room temperature.
Solution C:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of  Solution C: Add the NaHCO 3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Gas with 80% N 2 + 20% CO 2 to remove residual O 2 .
Solution D:
Composition per 10.0mL:
Sodium butyrate............................................................................0.7g
Sodium caproate ...........................................................................0.3g
Sodium octanoate........................................................................0.15g
Preparation of  Solution D: Add components to distilled/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 tempera-
ture.
Solution E:
Composition per 10.0mL:
Yeast extract..................................................................................1.0g
Thiamine·HCl .........................................................................100.0μg
p-Aminobenzoic acid................................................................40.0μg
D(+)-Biotin ...............................................................................10.0μg
Preparation of  Solution E: Add components to distilled/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.
Solution F:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.4g
Preparation of  Solution F: Add Na 2 S·9H 2 O to distilled/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 tempera-
ture.
Preparation of Medium: To 869.0mL of sterile cooled Solution A,
aseptically add the remaining sterile solutions in the following order:
solution B, solution C, solution D, solution E, and solution F. Mix thor-
oughly. Adjust pH to 7.7. Anaerobically distribute under 90% N 2 +
10% CO 2 into sterile tubes or flasks.
Use: For the cultivation of Acetivibrio ethanolgignens and Desulfovi-
brio sapovorans.