Chlamydia Isolation Medium

[所属分类:培养基配方] [发布时间:2020-4-17] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Chlamydia Isolation Medium

培养基配方
Composition per 500.0mL:
Eagle minimum essential medium
with Earle salts, 10X......................................50.0mL
Fetal calf serum...........................................50.0mL
Selective supplement.......................................10.0mL
L- Glutamine solution.......................................5.0mL
pH 7.4 ± 0.2 at 25°C
Eagle Minimum Essential Medium with Earle Salts, 10X:
Composition per liter:
NaCl.........................................................6.8g
Glucose......................................................1.0g
KCl..........................................................0.4g
CaCl2 ·2H2O ................................................0.2g
MgCl2 ·6H2O ................................................0.2g
NaH2PO4 ....................................................0.15g
L- Arginine..................................................0.1g
L- Lysine...................................................0.06g
L- Isoleucine...............................................0.05g
L- Leucine .................................................0.05g
L- Threonine................................................0.05g
L- Valine...................................................0.05g
L- Tyrosine ................................................0.04g
L- Phenylalanine............................................0.03g
L- Histidine................................................0.03g
L- Cystine..................................................0.02g
L- Methionine...............................................0.02g
L- Tryptophan...............................................0.01g
i-Inositol..................................................2.0mg
Calcium pantothenate .......................................1.0mg
Choline chloride............................................1.0mg
Folic acid .................................................1.0mg
Nicotinamide................................................1.0mg
Pyridoxal...................................................1.0mg
Thiamine·HCl ..............................................1.0mg
Riboflavin..................................................0.1mg
Preparation of Eagle Minimum Essential Medium with
Earle Salts, 10X: Add components to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.4 with 7.5%
Na2CO3 solution. Filter sterilize.
Selective Supplement:
Composition per 10.0mL:
Glucose.....................................................0.594g
Vancomycin...................................................0.05g
Gentamicin...................................................0.01g
Amphotericin B ..............................................2.0mg
Cycloheximide................................................2.0mg
Preparation of Selective Supplement: Add components to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Glutamine Solution:
Composition per 100.0mL:
L- Glutamine ................................................2.92g
NaCl (0.85% solution)......................................100.0mL
Preparation of Glutamine Solution: Add the glutamine to the
0.85% NaCl solution. Mix thoroughly. Filter sterilize.
Preparation of Medium: Aseptically combine 50.0mL of sterile
Eagle minimum essential medium with Earle salts, 10X, 50.0mL of fe-
tal calf serum, 10.0mL of selective supplement, and 5.0mL of sterile
glutamine solution. Bring volume to 500.0mL with sterile distilled/de-
ionized water. Mix thoroughly. Aseptically distribute into sterile tubes
or flasks.
Use: For the isolation and cultivation of Chlamydia species.