Anaeromyxobacter Medium (DSMZ Medium 1200)

[所属分类:培养基配方] [发布时间:2019-10-17] [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

Anaeromyxobacter Medium
(DSMZ Medium 1200)
Composition per liter:
Solution A..............................................................................900.0mL
Solution B................................................................................90.0mL
Solution C................................................................................18.0mL
Solution D................................................................................18.0mL
Solution F.................................................................................18.0mL
Solution E ..................................................................................4.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per 900.0mL:
NaCl..............................................................................................1.0g
MgCl2 ·6H2O..................................................................................0.5g
Sodium acetate..............................................................................0.4g
NH4Cl ...........................................................................................0.3g
KCl................................................................................................0.3g
KH2PO4 .........................................................................................0.2g
CaCl2 ·2H2O..............................................................................15.0mg
Resazurin ...................................................................................1.0mg
Selenite/tungstate solution.........................................................2.0mL
Trace element solution SL-10B.................................................1.0mL
Selenite/Tungstate Solution:
Composition per liter:
NaOH............................................................................................0.5g
Na2WO4 ·2H2O...........................................................................4.0mg
Na2SeO3 ·5H2O...........................................................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution SL-10B:
Composition per liter:
FeCl2 ·4H2O...................................................................................1.5g
H3BO3 ....................................................................................300.0mg
CoCl2 ·6H2O ...........................................................................190.0mg
MnCl2 ·4H2O...........................................................................100.0mg
ZnCl2 ........................................................................................70.0mg
Na2MoO 4 ·2H2O .......................................................................36.0mg
NiCl2 ·6H2O..............................................................................24.0mg
CuCl2 ·2H2O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10B:  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 compo-
nents. Mix thoroughly.
Preparation of Solution A:  Add components to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Sparge with 20%
CO 2 + 80% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Solution B:
Composition per 100.0mL:
NaHCO3 ........................................................................................5.0g
Preparation of Solution B:  Add NaHCO 3  to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with 20%
CO 2 + 80% H 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Solution C:
Composition per 50.0mL:
DL -Dithiothreitol....................................................................385.0mg
Preparation of Solution C:  Add  DL -dithiothreitol to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Solution D:
Composition per 50.0mL:
L -Cysteine·HCl·H 2 O................................................................37.5mg
Na 2 S·9H 2 O solution.................................................................10.0mL
Preparation of Solution D:  Add  L -cysteine·HCl·H 2 O to distilled/
deionized water and bring volume to 40.0mL. Mix thoroughly. Add
10.0mL Na 2 S·9H 2 O solution. Sparge with 20% CO 2 + 80% H 2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na2S·9H2O...............................................................................40.0mg
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.
Solution E:
Composition per 50.0mL:
Fumarate.......................................................................................4.0g
Preparation of Solution E:  Add fumarate to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100%
N 2 . Filter sterilize.
Solution F (Vitamin Solution):
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H2O..................................................................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 Solution F (Vitamin Solution):  Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Aseptically and anoxically add 1.0mL
solution B per 10 mL solution A. Adjust pH to 7.2. Add 0.2mL solution
C, 0.2mL solution D, 0.2mL solution F, and 0.05mL solution E, each
per 10mL solution A.
Use: For the cultivation and maintenance of Anaeromyxobacter spp.