Methanococcus Medium

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanococcus Medium

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

Composition per liter:

NaCl............................................................................................18.0g
Mg 2 SO 4 ·7H 2 O.............................................................................3.45g
MgCl 2 ·2H 2 O................................................................................2.75g
Pancreatic digest of casein............................................................2.0g
Yeast extract..................................................................................2.0g
Sodium acetate..............................................................................1.0g
L -Cysteine·HCl..............................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
NH 4 HCO 3 .....................................................................................0.5g
KCl............................................................................................0.335g
NH 4 Cl .......................................................................................0.225g
CaCl 2 ·2H 2 O.................................................................................0.14g
KH 2 PO 4 .......................................................................................0.14g
Calcium  DL -pantothenate...........................................................5.0mg
Na 2 SeO 3 .....................................................................................2.0mg
FeSO 4 ·2H 2 O...............................................................................1.0mg
Resazurin ...................................................................................1.0mg
Trace minerals stock solution .....................................................10mL
pH 6.5 ± 0.2 at 25°C
Trace Minerals Stock Solution:
Composition per liter:
Nitrilotriacetic acid.......................................................................1.5g
MnSO 4 ·H 2 O..................................................................................0.5g
CoCl 2 ............................................................................................0.1g
ZnSO 4 ...........................................................................................0.1g
AIK(SO 4 ) 2 ·12H 2 O ......................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
NiCl 2 ...........................................................................................0.01g
Preparation of Trace Minerals Stock Solution: Add nitrilotri-
acetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5
with KOH. Add remaining components one at a time. Add distilled/de-
ionized water to 1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare medium anaerobically under
80% N 2 + 20% CO 2 . Add components, except Na 2 CO 3 ,  L -
cysteine·HCl, and Na 2 S·9H 2 O, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool
while sparging with 80% N 2 + 20% CO 2 . Add Na 2 CO 3 ,  L -
cysteine·HCl, and Na 2 S·9H 2 O. Dispense into tubes, bottles, or flasks
under an atmosphere of 80% H 2 + 20% CO 2 . Seal with butyl rubber
stoppers secured with aluminum crimp seals. Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature. Pressurize the head
space to 69kPA with 80% H 2 + 20% CO 2 .
Use:  For the cultivation of Methanococcus species.