MG Medium

[所属分类:培养基配方] [发布时间:2021-5-27] [发布人:网站管理员2] [阅读次数:] [返回]

MG 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..........................................................................................100.0g
MgSO 4 ·7H 2 O..............................................................................3.45g
MgCl 2 ·6H 2 O................................................................................2.75g
Sodium acetate..............................................................................1.0g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O ................................................................................0.14g
K 2 HPO 4 ·3H 2 O ............................................................................0.14g
Resazurin...................................................................................1.0mg
NaHCO 3  solution.....................................................................80.0mL
Trimethylamine·HCl solution..................................................20.0mL
Na 2 CO 3 solution......................................................................10.0mL
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
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure–121°C.
Trimethylamine·HCl Solution:
Composition per 20.0mL:
Trimethylamine·HCl.....................................................................5.0g
Preparation of Trimethylamine·HCl Solution:  Add trimethyl-
amine·HCl to distilled/deionized water and bring volume to 20.0mL.
Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C.
Na 2 CO 3 Solution:
Composition per 10.0mL:
Na 2 CO 3  .........................................................................................0.5g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3  to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°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 approximately 500.0mL 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 100% N 2 .
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................................0.5g
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....................................................................................0.5g
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: Add components, except NaHCO 3  solu-
tion, trimethylamine·HCl solution, Na 2 CO 3 solution, vitamin solution,
L -cysteine·HCl solution, and Na 2 S·9H 2 O solution, to distilled/deion-
ized water and bring volume to 860.0mL. Mix thoroughly. Sparge with
100% N 2 for 20 min. Then sparge with 80% N 2 + 20% CO 2 for 10 min.
Anaerobically distribute into tubes or bottles. Autoclave for 15 min at
15 psi pressure–121°C. Aseptically and anaerobically add 80.0mL of
sterile NaHCO 3  solution, 20.0mL of sterile trimethylamine·HCl solu-
tion, 10.0mL of sterile Na 2 CO 3 solution, 10.0mL of sterile vitamin so-
lution, 10.0mL of sterile  L -cysteine·HCl solution, and 10.0mL of sterile
Na 2 S·9H 2 O solution. Mix thoroughly.
Use: For the cultivation and maintenance of Methanohalobium spe-
cies, Methanohalophilus halophilus, and Methanohalophilus species.