Methanosarcina Medium (BCYT)

[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 Medium
(BCYT)

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

Composition per liter:

KHCO 3 ..........................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
NaCl..............................................................................................0.6g
Pancreatic digest of casein............................................................0.5g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 ·2H 2 O.................................................................................0.08g
Resazurin ...................................................................................1.0mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine·HCl·H 2 O solution ..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Methanol....................................................................................5.0mL
pH 6.8 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.8g
FeCl 3 ·6H 2 O.................................................................................1.35g
NaCl..............................................................................................1.0g
NiCl 2 ·6H 2 O.................................................................................0.12g
MnCl 2 ·4H 2 O..................................................................................0.1g
CaCl 2 ·2H 2 O...................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
Na 2 SeO 3 ·5H 2 O..........................................................................0.026g
CuCl 2 ·2H 2 O ..............................................................................0.025g
CoCl 2 ·6H 2 O ..............................................................................0.024g
Na 2 MoO 4 ·2H 2 O ........................................................................0.024g
H 3 BO 3 .........................................................................................0.01g
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to approximately 500.0mL of distilled/deionized water. Dissolve
by adding KOH and adjust pH to 6.5. Add remaining components.
Bring volume to 1.0L with additional distilled/deionized water. Adjust
pH to 7.0 with KOH.
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/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-

ilize. Sparge with 80% N 2 + 20% CO 2 .

L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O...................................................................0.25g
Preparation of  L -Cysteine·HCl·H 2 O Solution: Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Autoclave under 80% N 2 + 20% CO 2 for 15 min at 15
psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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 10.0mL. Mix thoroughly.
Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: Add components, except vitamin solu-
tion,  L -cysteine·HCl·H 2 O solution, and Na 2 S·9H 2 O solution, to dis-
tilled/deionized water and bring volume to 960.0mL. Mix thoroughly.
Sparge under 80% N 2 + 20% CO 2 for 3–4 min. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of
sterile vitamin solution, 10.0mL of sterile  L -cysteine·HCl·H 2 O solu-
tion, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly.
Aseptically and anaerobically distribute into sterile screw-cappped
bottles under 80% N 2 + 20% CO 2 .
Use: For the cultivation and maintenance of Methanosarcina mazei
and Methanosarcina thermophila.