Modified VWM Medium (DSMZ Medium 503a)

[所属分类:培养基配方] [发布时间:2021-6-4] [发布人:网站管理员2] [阅读次数:] [返回]

Modified VWM Medium
(DSMZ Medium 503a)

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

Composition per 1013.4mL:

Solution A..............................................................................940.0mL
Solution E ................................................................................50.0mL
Solution F.................................................................................10.0mL
Solution G................................................................................10.0mL
Solution B..................................................................................1.0mL
Solution C..................................................................................1.0mL
Solution D..................................................................................1.0mL
Solution H..................................................................................0.4mL
pH 7.3 ± 0.2 at 25°C
Solution A:
Composition per 940.0mL:
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin...................................................................................0.5mg
Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas
atmosphere. Add components to distilled/deionized water and bring vol-
ume to 940.0mL. 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. Cool to
25°C.
Solution B:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Solution B: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so-
lution. Mix thoroughly. Add distilled/deionized water and bring vol-
ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with
80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 25°C.
Solution C:
Composition per liter:
Pyridoxine hydrochloride......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Solution C: Add components to distilled/deionized
water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough-
ly. Filter sterilize.
Solution D:
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Solution D: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%
N 2 . Filter sterilize.
Solution E:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of Solution E:  Add NaHCO 3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 25°C.
Solution F:
Composition per 10.0mL:
Taurine ..........................................................................................2.5g
Preparation of Solution F:  Add taurine to distilled/deionized wa-
ter and bring volume to 100.0mL. Mix thoroughly. Sparge with 100%
N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C.
Solution G :
Composition per 10.0mL:
Na 2 S·9H 2 O................................................................................0.125g
Preparation of Solution G:  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 25°C.
Solution H :
Composition per 10.0mL:
Na-dithionite.................................................................................0.5g
Preparation of Solution H:  Add Na-dithionite to distilled/deion-
ized 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 25°C.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B,
1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so-
lution F, 10.0mL solution G, and 0.4mL solution H to 940.0mL solution
A. Distribute anaerobically under 80% N 2  + 20% CO 2 into appropriate
vessels. The pH should be 7.2–7.4.
Use: For the cultivation of Desulfonispora thiosulfatigenes.