Methanosarcina frisia Medium

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanosarcina frisia 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
NaHCO 3 ........................................................................................5.0g
MgCl 2 ·6H 2 O..................................................................................4.0g
MgSO 4 ·7H 2 O............................................................................. 3.45g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................2.0g
Sodium acetate..............................................................................1.0g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O.................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
L -Cysteine·HCl..............................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
Fe(NH 4 ) 2 (SO 4 ) 2 ·7H 2 O ...............................................................2.0mg
Resazurin ...................................................................................1.0mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Methanol solution......................................................................2.0mL
pH 6.8–7.0 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O............................................................................... 3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O............................................................................... 0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ......................................................................... 0.01g
NiCl 2 ·6H 2 O...............................................................................0.025g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Adjust pH to 7.0. Add distilled/de-
ionized water to 1.0L.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg

Vitamin B 12 ................................................................................0.1mg

Preparation of Vitamin Solution : Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80%
H 2 + 20% CO 2 .
Methanol Solution:
Composition per 10.0mL:
Methanol..................................................................................10.0mL
Preparation of Methanol Solution:  Sparge 10.0mL of methanol
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except methanol solution, to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. An-
aerobically distribute into tubes or flasks fitted with butyl rubber
stoppers. Autoclave for 15 min at 15 psi pressure–121°C. Anaerobical-
ly add 10.0mL of sterile methanol solution to each liter of medium or,
using a syringe, inject the appropriate amount of sterile methanol solu-
tion into individual tubes containing medium.
Use:  For the cultivation and maintenance of Methanosarcina frisia.