Hydroxybenzoate Medium

[所属分类:培养基配方] [发布时间:2021-4-19] [发布人:李佳颐] [阅读次数:] [返回]

Hydroxybenzoate Medium

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

Composition per 1002.0mL:

Solution A..............................................................................920.0mL
Solution B................................................................................50.0mL
Solution E (Vitamin solution)..................................................10.0mL
Solution F.................................................................................10.0mL
Solution G................................................................................10.0mL
Solution C (Trace elements solution SL-10) .............................1.0mL
Solution D..................................................................................1.0mL
pH 7.2–7.5 at 25°C
Solution A:
Composition per 920.0mL:
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 920.0mL. Mix thoroughly. Sparge with 80%
N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 50.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of Solution B:  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.
Solution 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 Solution C (Trace Elements Solution SL-10):
Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Gas under 80% N 2  + 20% CO 2 . Autoclave for
15 min at 15 psi pressure–121°C.
Solution D:
Composition per 10.0mL:
NaOH.........................................................................................5.0mg
Na 2 WO 4 ·2H 2 O..........................................................................40.0μg
Na 2 SeO 3 ·5H 2 O..........................................................................30.0μg
Preparation of Solution D:  Add components 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.
Solution E (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 Solution E (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 pres-
sure–121°C.
Solution F:
Composition per 10.0mL:
Sodium dihydroxybenzoate ..........................................................0.4g
Preparation of Solution F:  Add sodium dihydroxybenzoate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize. Sparge with 80% N 2 + 20% CO 2 .
Solution G:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
Preparation of Solution G:  Add Na 2 S·9H 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.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Aseptically and anaerobically combine 920.0mL of sterile
solution A, 50.0mL of sterile solution B, 1.0mL of sterile solution C,
1.0mL of sterile solution D, 10.0mL of sterile solution E, 10.0mL of sterile
solution F, and 10.0mL of sterile solution G. Mix thoroughly. Aseptically
and anaerobically distribute into sterile tubes or flasks.
Use: For the cultivation and maintenance of Clostridium species.