Heterotrophic Agar H3P

[所属分类:培养基配方] [发布时间:2021-4-6] [发布人: ] [阅读次数:] [返回]

Heterotrophic Agar H3P

培养基配方

Composition per 1010.0mL:
Solution B..............................................................................855.0mL
Solution A................................................................................50.0mL
Solution E ................................................................................50.0mL
Solution C................................................................................20.0mL
Solution D................................................................................30.0mL
Standard vitamin solution..........................................................5.0mL
pH 6.8 ± 0.2 at 25°C
Solution A:
Composition per 50.0mL:
Na 2 HPO 4 ·2H 2 O.............................................................................2.9g
KH 2 PO 4 .........................................................................................2.3g
Preparation of Solution A:  Add components to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.
Solution B:
Composition per 855.0mL:
Agar ............................................................................................15.0g
NH 4 Cl ...........................................................................................1.0g
MgSO 4 ·7H 2 O................................................................................0.5g
CaCl 2 ·2H 2 O.................................................................................0.01g
MnCl 2 ·4H 2 O...............................................................................5.0mg
NaVO 3 ·H 2 O ...............................................................................5.0mg
Trace elements solution SL-6 ....................................................5.0mL
Trace Elements Solution SL-6:
Composition per liter:
MnCl 2 ·4H 2 O..................................................................................0.5g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O ..................................................................................0.2g
ZnSO 4 ·7H 2 O................................................................................ 0.1g
Na 2 MoO 4 ·2H 2 O ......................................................................... 0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2 ·2H 2 O ................................................................................0.01g
Preparation of Trace Elements Solution SL-6:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Solution B: Add components to distilled/deionized water and bring volume to 855.0mL. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C.
Solution C:
Composition per 20.0mL:
Ferric ammonium citrate.............................................................0.05g
Preparation of Solution C:  Add ferric ammonium citrate to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.
Solution D:
Composition per 30.0mL:
Disodium succinate.......................................................................1.0g
DL -Malate......................................................................................1.0g
Sodium acetate..............................................................................1.0g
Yeast extract..................................................................................1.0g
Preparation of Solution D:  Add components to distilled/deionized water and bring volume to 30.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.
Solution E:
Composition per 50.0mL:
D -Glucose......................................................................................2.0g
D -Mannitol....................................................................................1.0g
Sodium lactate...............................................................................1.0g
Sodium pyruvate...........................................................................1.0g
Preparation of Solution E:  Add components to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize. Warm to 50°C.
Standard Vitamin Solution:
Composition per 100.0mL:
Calcium  DL -pantothenate.........................................................50.0mg
Nicotinic acid...........................................................................50.0mg
Pyridoxine HCl........................................................................50.0mg
Thiamine..................................................................................50.0mg
Riboflavin ................................................................................10.0mg
B 12 ..............................................................................................1.0mg
Folic acid....................................................................................0.2mg
Biotin .........................................................................................0.1mg
Preparation of Standard Vitamin Solution:  Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Warm to 50°C.
Preparation of Medium: Aseptically combine 50.0mL of sterile solution A, 855.0mL of sterile solution B, 20.0mL of sterile solution C, 30.0mL of sterile solution D, 50.0mL of sterile solution E, and 5.0mL of sterile standard vitamin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use:  For the heterotrophic growth, isolation, and purity checking of a wide variety of aerobic bacteria, including Ancylobacter species, Aquaspirillum species, Azospirillum species, Beijerinckia species, Derxia gummosa, Herbaspirillum seropedicae, Rhodococcus species, Xanthobacter species, and Xanthomonas campestris.