Mycoplasma Medium, Revised

[所属分类:培养基配方] [发布时间:2021-6-16] [发布人:网站管理员2] [阅读次数:] [返回]

Mycoplasma Medium, Revised

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

Composition per 1030.0mL:

Noble agar...................................................................................10.0g
Distilled water........................................................................360.0mL
Heart infusion broth...............................................................300.0mL
Pig serum, heat inactivated....................................................200.0mL
Enzymatic hydrolysate of lactalbumin..................................100.0mL
Hanks’ balanced salt solution, 10X.........................................40.0mL
Fresh yeast extract solution .....................................................20.0mL
Phenol Red (0.25% solution)...................................................10.0mL
Heart Infusion Broth:
Composition per liter:
Beef heart, infusion from..........................................................500.0g
Tryptose......................................................................................10.0g
NaCl..............................................................................................5.0g
Preparation of Heart Infusion Broth: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gen-
tly heat and bring to boiling.
Enzymatic Hydrolysate of Lactalbumin:
Composition per 100.0mL:
Enzymatic hydrolysate of lactalbumin.........................................5.0g
Phosphate buffered saline, 1X, pH7.0...................................100.0mL
Preparation of Enzymatic Hydrolysate of Lactalbumin: Add
enzymatic hydrolysate of lactalbumin to 100.0mL of phosphate buff-
ered saline, 1X, pH 7.0.
Phosphate Buffered Saline Solution, 1X:
Composition per liter:
NaCl..............................................................................................8.0g
Na 2 HPO 4 ·7H 2 O.............................................................................2.16
KCl................................................................................................0.2g
KH 2 PO 4 .........................................................................................0.2g
MgCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
Preparation of Phosphate Buffered Saline Solution, 1X: Add
components to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly.
Hanks’ Balanced Salt Solution, 10X:
Composition per liter:
Na 2 Cl...........................................................................................80.0g
Glucose .......................................................................................10.0g
KCl................................................................................................4.0g
CaCl 2 .............................................................................................1.4g
MgCl 2 ·6H 2 O..................................................................................1.0g
MgSO 4 ·7H 2 O................................................................................1.0g
Na 2 HPO 4 ·7H 2 O.............................................................................0.9g
KH 2 PO 4 .........................................................................................0.6g
Preparation of Hanks’ Balanced Salt Solution, 10X: Add
components to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly.
Fresh Yeast Extract Solution:
Composition per 100.0mL:
Baker’s yeast, live, pressed, starch-free......................................25.0g
Preparation of Fresh Yeast Extract Solution : Add the live Bak-
er’s yeast to 100.0mL of distilled/deionized water. Autoclave for 90
min at 15 psi pressure–121°C. Allow to stand. Remove supernatant so-
lution. Adjust pH to 6.6–6.8.
Preparation of Medium: Add agar and heart infusion broth to dis-
tilled/deionized water and bring volume to 660.0mL. Mix thoroughly. Ad-
just pH to 7.4. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
add pig serum, enzymatic hydrolysate of lactalbumin, Hanks’ balanced
salt solution, 10X, fresh yeast extract solution, and Phenol Red solution.
Mix thoroughly. Aseptically distribute into sterile tubes.
Use: For the cultivation and maintenance of Mycoplasma dispar,
Mycoplasma flocculare, and Mycoplasma hyopneumoniae.