GÖ1 Medium

[所属分类:培养基配方] [发布时间:2021-3-18] [发布人: ] [阅读次数:] [返回]

GÖ1 Medium

培养基配方

Composition per liter:
Sucrose........................................................................................17.1g
Sodium acetate............................................................................10.0g
NaCl............................................................................................2.25g
Pancreatic digest of casein............................................................2.0g
Yeast extract..................................................................................2.0g
NaHCO 3 ......................................................................................0.85g
MgSO 4 ·7H 2 O................................................................................0.5g
NH 4 Cl ...........................................................................................0.5g
K 2 HPO 4 ....................................................................................0.348g
CaCl 2 ·2H 2 O................................................................................0.25g
KH 2 PO 4 ....................................................................................0.227g
FeSO 4 ·7H 2 O ..............................................................................2.0mg
Resazurin ...................................................................................1.0mg
NaHCO 3 solution.....................................................................40.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine·HCl·H 2 O solution..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 6.5–6.8 at 25°C
NaHCO 3 Solution:
Composition per 50.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
DL -Calcium 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 sterilize. 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.....................................................................0.3g
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 100% N 2 for 15 min at 15 psi pres-
sure–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 distilled/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. Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 100% N 2 .
Preparation of Medium: Add components, except NaHCO 3 solution, vitamin solution,  L -cysteine·HCl·H 2 O solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 920.0mL. 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 40.0mL of sterile NaHCO 3 solution, 20.0mL of sterile vitamin solution, 10.0mL of sterile  L -cysteine·HCl·H 2 O solution, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile screw-capped bottles under 80% N 2 + 20% CO 2 .
Use: For the cultivation of Methanosarcina mazei.