Magnetospirillum Semi-solid Medium (DSMZ Medium 380)

[所属分类:培养基配方] [发布时间:2021-5-12] [发布人:网站管理员2] [阅读次数:] [返回]

Magnetospirillum Semi-solid Medium
(DSMZ Medium 380)

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

Composition per liter:

Agar ..............................................................................................1.3g
KH 2 PO 4 .......................................................................................0.68g
L (+)-Tartaric acid........................................................................0.37g
Succinic acid...............................................................................0.37g
NaNO 3 ........................................................................................0.12g
Na-thioglycolate..........................................................................0.05g
Na-acetate ...................................................................................0.05g
Resazurin ...................................................................................0.5mg
Vitamin solution.......................................................................10.0mL
Trace elements solution .............................................................5.0mL
Ferric quinate solution...............................................................2.0mL
pH 6.8 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate .....................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Ferric Quinate Solution:
Composition per 100.0mL:
FeCl 3 ·6H 2 O.................................................................................0.45g
Quinic acid..................................................................................0.19g
Preparation of Ferric Quinate Solution:  Add components to
distilled/deionized water and bring volume to 100.0mL. Sparge with
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°C.
Preparation of Medium: Add components, except vitamin solu-
tion and ferric quinate solution, to distilled/deionized water and bring
volume to 988.0mL. Purge medium with N 2 gas for 10 min. Mix thor-
oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°C.
Aseptically and anerobically add 10.0mL vitamin solution and 2.0mL
ferric quinate solution. Mix thoroughly. Purge medium with N 2 gas for
10 min. Dispense 12mL of medium per 16 x 150mm anaerobe screw-
cap tube under N 2 gas. Prior to inoculation, remove caps briefly under
air, tighten the caps again, and wait several hours to establish oxygen
gradients. The medium should be slightly pink in color. Strongly re-
duced conditions will not support growth of the organism. During
growth O 2  will be consumed, resazurin decolorized, and the pH in-
creased. Feed oxygen (by adding air) and succinic acid from sterile
0.05M solution (to maintain pH below 7.0). If higher densities of mag-
netic cell are wanted, ferric quinate also can be fed. For transfer use cell
material which has been concentrated at the glass wall of the culture

vessel by means of a magnetic rod attached outside.

Use:  For the cultivation of Magnetospirillum magnetotacticum
(Aquaspirillum magnetotacticum) and Magnetospirillum gryphiswald-
ense.