Atopobium/Olsenella Medium (LMG Medium 152)

[所属分类:培养基配方] [发布时间:2019-11-14] [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

Atopobium/Olsenella Medium
(LMG Medium 152)
Composition per liter:
Yeast extract................................................................................10.0g
Peptone .........................................................................................5.0g
Casitone ........................................................................................5.0g
Glucose.........................................................................................5.0g
(NH4)2SO4 ....................................................................................0.5g
L -Cysteine·HCl .............................................................................0.5g
Resazurin ...................................................................................1.0mg
Mineral solution.......................................................................40.0mL
Fatty acid mixture......................................................................3.1mL
Tween™ 80................................................................................2.0mL
Hemin solution...........................................................................0.5mL
Vitamin K 1 .................................................................................0.2mL
pH 6.9 ± 0.2 at 25°C
Mineral Solution:
Composition per liter:
NaHCO3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K2HPO4 .........................................................................................1.0g
KH2PO4 .........................................................................................1.0g
MgSO4 ·7H2O..............................................................................0.48g
CaCl2 ·2H2O ..................................................................................0.3g
Preparation of Mineral Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Fatty Acid Mixture:
Composition per 31.0mL:
Acetic acid...............................................................................17.0mL
Propionic acid............................................................................6.0mL
n-Butyric acid ............................................................................4.0mL
n-Valeric acid.............................................................................1.0mL
iso-Valeric acid ..........................................................................1.0mL
iso-Butyric acid..........................................................................1.0mL
DL -2-Methylbutyric acid............................................................1.0mL
Preparation of Fatty Acid Mixture : Combine components. Mix
thoroughly. Adjust pH to 7.5 with concentrated NaOH.
Hemin Solution:
Composition per 1.0mL:
Hemin.........................................................................................5.0mg
NaOH (1N solution)...................................................................1.0mL
Preparation of Hemin Solution: Add hemin to 1.0mL of NaOH
solution. Mix thoroughly.
Preparation of Medium: Add components, except  L -cysteine·HCl,
hemin solution, and fatty acid mixture, to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boil-
ing. Continue boiling for 5 min. Cool to room temperature while sparg-
ing with 100% CO 2 . Add  L -cysteine·HCl, hemin solution, and fatty
acid mixture. Adjust pH to 6.9 with 8N NaOH while continuing to
sparge with 100% CO 2 . After pH has been reached, sparge with 100%
N 2 . Anaerobically distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure–121°C.
Use:  For the cultivation and maintenance of Atopobium rimae and
Olsenella uli.