Methanococcus deltae 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 deltae 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............................................................................................35.0g
NaHCO 3 ........................................................................................5.0g
MgCl 2 ·6H 2 O..................................................................................4.0g
NH 4 Cl ...........................................................................................2.7g
Sodium acetate..............................................................................2.5g
L -Cysteine·HCl..............................................................................0.3g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
Na 2 S·9H 2 O....................................................................................0.3g
MgSO 4 ·7H 2 O..............................................................................0.13g
Resazurin ...................................................................................1.0mg
(NH 4 ) 2 SO 4 ..................................................................................0.3mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine·HCl solution...........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.9 ± 0.2 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
NiCl 2 ·6H 2 O...............................................................................0.025g
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
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/
deionized water and bring volume to 1.0L. Adjust pH to 7.0. Mix thor-
oughly.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl .............................................................................0.3g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 100% N 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 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except  L -cysteine·HCl solution and
Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Anaerobically distribute into tubes or flasks
fitted with butyl rubber stoppers. Autoclave for 15 min at 15 psi pres-
sure–121°C. Anaerobically add 10.0mL of sterile  L -cysteine·HCl solu-
tion and 10.0mL of sterile Na 2 S·9H 2 O solution to each liter of medium
or, using a syringe, inject the appropriate amount of sterile  L -
cysteine·HCl solution and sterile Na 2 S·9H 2 O solution into individual
tubes containing medium.
Use:  For the cultivation and maintenance of Methanococcus deltae.