Mitsuokella dentalis Medium

[所属分类:培养基配方] [发布时间:2021-6-3] [发布人:网站管理员2] [阅读次数:] [返回]

Mitsuokella dentalis Medium

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

Composition per 1003.3mL:

Yeast extract................................................................................10.0g
Beef extract...................................................................................5.0g
Glucose.........................................................................................5.0g
Trypticase™..................................................................................5.0g
L -Cysteine·HCl .............................................................................0.5g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
Resazurin...................................................................................1.0mg
Bovine serum...........................................................................50.0mL
Mineral solution.......................................................................40.0mL
Hemin solution...........................................................................1.0mL
Vitamin K 1  solution....................................................................0.2mL
pH 6.9 ± 0.2 at 25°C
Bovine Serum:
Composition per 50.0mL:
Bovine serum...........................................................................50.0mL
Preparation of Bovine Serum:  Incubate 50.0mL of bovine serum
in a GasPak™ container overnight to make anaerobic.
Mineral Solution:
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
MgSO 4 ·7H 2 O..............................................................................0.48g
CaCl 2 ·2H 2 O ..................................................................................0.3g
Preparation of Mineral Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Hemin Solution:
Composition per 100.0mL:
Hemin ........................................................................................5.0mg
NaOH (0.002% solution)...........................................................1.0mL
Preparation of Hemin Solution: Add hemin to 1.0mL of NaOH
solution. Mix thoroughly.
Vitamin K 1 Solution:
Composition per 100.0mL:
Vitamin K 1 ..................................................................................1.09g
Preparation of Vitamin K 1 Solution:  Add vitamin K 1 to 99.0mL
of absolute ethanol. Store in the dark at 4°C.
Preparation of Medium: Add components, except  L -cysteine·HCl
and bovine serum, to distilled/deionized water and bring volume to
950.0mL. Mix thoroughly. Gently heat and bring to boiling. Continue
boiling for 5 min. Cool to room temperature while sparging with 100%
CO 2 . Add  L -cysteine·HCl. Mix thoroughly. Adjust pH to 6.0 with 8N
NaOH. After pH has been reached, change sparging gas to 100% N 2 .
Anaerobically distribute into bottles under 100% N 2 . Autoclave for 15
min at 15 psi pressure–121°C. Aseptically and anaerobically add
50.0mL of bovine serum. Mix thoroughly.
Use: For the cultivation and maintenance of Mitsuokella dentalis.