GC Agar with Chloramphenicol, Tetracycline, and Ampicillin

[所属分类:培养基配方] [发布时间:2021-3-9] [发布人: ] [阅读次数:] [返回]

GC Agar with Chloramphenicol,

Tetracycline, and Ampicillin

培养基配方

Composition per 1040.0mL:
GC agar base, 2X...................................................................500.0mL
Hemoglobin solution..............................................................500.0mL
Supplement solution ................................................................10.0mL
Ampicillin solution..................................................................10.0mL
Tetracycline solution................................................................10.0mL
Chloramphenicol solution........................................................10.0mL
pH 7.2 ± 0.2 at 25°C
GC Agar Base, 2X:
Composition per 500.0mL:
Agar ............................................................................................10.0g
Pancreatic digest of casein............................................................7.5g
Peptic digest of animal tissue........................................................7.5g
NaCl..............................................................................................5.0g
K 2 HPO 4 .........................................................................................4.0g
Cornstarch.....................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
Source:  GC agar base is available as a premixed powder from BD Diagnostic Systems. This base may be replaced by GC medium base available from BD Diagnostic Systems.
Preparation of GC Agar Base, 2X: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Hemoglobin Solution:
Composition per 500.0mL:
Bovine hemoglobin.....................................................................10.0g
Preparation of Hemoglobin Solution: Add bovine hemoglobin to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C.
Supplement Solution:
Composition per liter:
Glucose.....................................................................................100.0g
L- Cysteine·HCl............................................................................25.9g
L- Glutamine ................................................................................10.0g
L- Cystine.......................................................................................1.1g
Adenine.........................................................................................1.0g
Nicotinamide adenine dinucleotide ............................................0.25g
Vitamin B 12 ...................................................................................0.1g
Thiamine pyrophosphate ..............................................................0.1g
Guanine·HCl...............................................................................0.03g
Fe(NO 3 ) 3 ·6H 2 O...........................................................................0.02g
p-Aminobenzoic acid................................................................0.013g
Thiamine·HCl ............................................................................3.0mg
Source: The supplement solution (IsoVitaleX ® enrichment) is available from BD Diagnostic Systems. This enrichment may be replaced by supplement VX from BD Diagnostic Systems.
Preparation of Supplement Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Ampicillin Solution:
Composition per 10.0mL:
Ampicillin...................................................................................0.01g
Preparation of Ampicillin Solution: Add ampicillin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Tetracycline Solution:
Composition per 10.0mL:
Tetracycline................................................................................5.0mg
Preparation of Tetracycline Solution: Add tetracycline to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Chloramphenicol Solution:
Composition per 10.0mL:
Chloramphenicol........................................................................5.0mg
Preparation of Chloramphenicol Solution: Add chloramphenicol to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 500.0mL of sterile GC agar base, aseptically add 500.0mL of sterile hemoglobin solution at 45°–50°C. Mix thoroughly. Aseptically add 10.0mL of sterile supplement solution, 10.0mL of sterile ampicillin solution, 10.0mL of sterile chloramphenicol solution, and 10.0mL of sterile tetracycline solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use:  For the cultivation and maintenance of Haemophilus parainfluenzae.