Anaerolinea Medium without Glucose (DSMZ Medium 1004)

[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

Anaerolinea Medium without Glucose
(DSMZ Medium 1004)
Composition per liter:
NaHCO3 ........................................................................................2.5g
NH4Cl .........................................................................................0.54g
MgCl2 ·6H2O..................................................................................0.2g
CaCl2 ·2H2O.................................................................................0.15g
KH2PO4 .......................................................................................0.14g
Yeast extract..................................................................................0.1g
Resazurin ...................................................................................1.0mg
L -Cysteine solution..................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Na2S·9H2O solution.................................................................10.0mL
Selenite/tungstate solution.........................................................1.0mL
Trace elements solution SL-11 ..................................................1.0mL
pH 7.0 ± 0.1 at 25°C
Na2S·9H2O Solution :
Composition per 100.0mL:
Na2S·9H2O..................................................................................0.25g
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 100.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
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 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.
L -Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O...................................................................0.25g
Preparation of  L -Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Trace Elements Solution SL-11:
Composition per liter:
FeCl2 ·4H2O...................................................................................1.5g
CoCl2 ·6H2O...........................................................................190.0mg
MnCl2 ·4H2O ..........................................................................100.0mg
ZnCl2· .......................................................................................70.0mg
Na2MoO4 ·H2O.........................................................................36.0mg
NiCl 2 ·6H2O..............................................................................24.0mg
H3BO3 ........................................................................................6.0mg
Na2-EDTA ....................................................................................5.2g
CuCl2· ·2H2O...............................................................................2.0mg
Preparation of Trace Elements Solution SL-11: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 6.0.
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.
Preparation of Medium: Add components, except vitamin solu-
tion, NaHCO 3 ,  L -cysteine solution, and Na 2 S·9H 2 O solution , to dis-
tilled/deionized water and bring volume to 970.0mL. Mix thoroughly.
Gently heat and bring to boiling. Cool to room temperature while
sparging with 20% CO 2 + 80% N 2 . Add solid bicarbonate. Mix thor-
oughly. Adjust pH to 7.0. Dispense into tubes or bottles. Autoclave for
15 min at 15 psi pressure–121°C. Cool to 25°C under 20% CO 2 + 80%
N 2 . Aseptically and anaerobically add sterile  L -cysteine, vitamin, and
Na 2 S·9H 2 O solutions. The final pH should be 7.0.
Use: For the cultivation and maintenance of Leptolinea tardivitalis.