GC Medium with Chloramphenicol

[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 Medium with Chloramphenicol

培养基配方

Composition per 1020.0mL:
GC agar base, 2X...................................................................500.0mL
Hemoglobin solution .............................................................500.0mL
Supplement solution................................................................10.0mL
Chloramphenicol solution........................................................10.0mL
pH 7.2 ± 0.2 at 25°C
GC Agar Base, 2 X :
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, 2 X : 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.
Chloramphenicol Solution:
Composition per 10.0mL:
Chloramphenicol........................................................................8.0mg
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 and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use: For the cultivation of Neisseria gonorrhoeae.