GV Medium

[所属分类:培养基配方] [发布时间:2021-3-22] [发布人: ] [阅读次数:] [返回]

GV Medium

培养基配方

Composition per 1051.0mL:

NaHCO 3 ........................................................................................3.0g
NaCl............................................................................................2.25g
Yeast extract..................................................................................1.0g
NH 4 Cl ...........................................................................................0.5g
K 2 HPO 4 .....................................................................................0.348g
KH 2 PO 4 .....................................................................................0.227g
FeSO 4 ·7H 2 O...............................................................................2.0mg
Resazurin ...................................................................................0.5mg
Vitamin solution.......................................................................10.0mL
Substrate solution.....................................................................10.0mL
L -Cysteine·HCl solution...........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
MgSO 4 ·7H 2 O solution.............................................................10.0mL

CaCl 2 ·2H 2 O solution................................................................10.0mL

Selenite-tungstate solution.........................................................1.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0 ± 0.2 at 25°C
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. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Substrate Solution:
Composition per 10.0mL:
Sodium crotonate.....................................................................10.0mg
Preparation of Substrate Solution: Add sodium crotonate to distilled/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 pressure–121°C.
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 thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave 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 distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Gas under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
MgSO 4 ·7H 2 O Solution:
Composition per 10.0mL:
MgSO 4 ·7H 2 O................................................................................0.5g
Preparation of MgSO 4 ·7H 2 O Solution: Add MgSO 4 ·7H 2 O to distilled/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 pressure–121°C.
CaCl 2 ·2H 2 O Solution:
Composition per 10.0mL:
CaCl 2 ·2H 2 O ................................................................................0.25g
Preparation of CaCl 2 ·2H 2 O Solution: Add CaCl 2 ·2H 2 O to distilled/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 pressure–121°C.
Selenite-Tungstate Solution:
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O..............................................................................4mg
Na 2 SeO 3 ·5H 2 O..............................................................................3mg
Preparation of Selenite-Tungstate Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.Sparge with 80% N 2 + 20% CO 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. Gas under 80% N 2  + 20% CO 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 substrate solution, MgSO 4 ·7H 2 O solution, CaCl 2 ·2H 2 O solution, selenite-tungstate solution,  L -cysteine·HCl solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL of sterile MgSO 4 ·7H 2 O solution, 10.0mL of sterile substrate solution, 10.0mL of sterile CaCl 2 ·2H 2 O solution, 1.0mL of sterile selenite-tungstate solution, 10.0mL of sterile L -cysteine·HCl solution, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Adjust pH to 7.0 to 7.2. Aseptically and anaerobically distribute into sterile tubes or flasks.
Use: For the cultivation and maintenance of Ilyobacter delafieldii.