Anaerolinea Medium (DSMZ Medium 1004)

[所属分类:培养基配方] [发布时间:2019-10-16] [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
(DSMZ Medium 1004)
Composition per liter:
NaHCO3 ........................................................................................2.5g
Yeast extract..................................................................................2.3g
NH4Cl .........................................................................................0.54g
MgCl2 ·6H2O .................................................................................0.2g
CaCl2 ·2H2O ................................................................................0.15g
KH2PO4 .......................................................................................0.14g
Resazurin...................................................................................1.0mg
Glucose solution......................................................................10.0mL
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.
Glucose Solution:
Composition per 10.0mL:
Glucose .........................................................................................2.2g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 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
NiCl2·6H2O..............................................................................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, Na 2 S·9H 2 O solution and glucose
solution, to distilled/deionized water and bring volume to 960.0mL.
Mix thoroughly. Gently heat and bring to boiling. Cool to room tem-
perature while sparging with 20% CO 2 + 80% N 2 . Add solid bicarbon-
ate. Mix thoroughly. 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 glucose,
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 Anaerolinea spp.