Minimum Essential Medium with Bicarbonate, Serum, and Antibiotics

[所属分类:培养基配方] [发布时间:2021-6-3] [发布人:网站管理员2] [阅读次数:] [返回]

Minimum Essential Medium with
Bicarbonate, Serum, and Antibiotics

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

Composition per 1100.0mL:

Minimum essential medium ..................................................950.0mL
Fetal bovine serum, heat inactivated .....................................100.0mL
NaHCO 3 solution.....................................................................40.0mL
Penicillin-streptomycin solution..............................................10.0mL
pH 7.2 ± 0.2 at 25°C
Minimum Essential Medium (MEM):
Composition per liter:
Inorganic salt solution............................................................400.0mL
Other component solution......................................................400.0mL
Amino acid solution...............................................................100.0mL
Vitamin solution.....................................................................100.0mL
Inorganic Salt Solution:
Composition per 400.0mL:
NaCl..............................................................................................6.8g
KCl................................................................................................0.4g
CaCl 2 , anhydrous..........................................................................0.2g
NaH 2 PO 4 ·H 2 O.............................................................................0.14g
MgSO 4 , anhydrous.................................................................97.67mg
Preparation of Inorganic Salt Solution : Add components to dis-
tilled/deionized water and bring volume to 400.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Other Component Solution:
Composition per 400.0mL:
D -Glucose......................................................................................1.0g
Phenol Red...............................................................................10.0mg
Preparation of Other Component Solution: Add components
to distilled/deionized water and bring volume to 400.0mL. Mix thor-
oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Amino Acid Solution:
Composition per 100.0mL:
L -Glutamine...........................................................................292.0mg
L -Arginine·HCl ......................................................................126.1mg
L -Lysine·HCl............................................................................72.5mg
L -Isoleucine..............................................................................52.0mg
L -Leucine .................................................................................52.0mg
L -Tyrosine, disodium salt.........................................................52.0mg
L -Threonine..............................................................................48.0mg
L -Valine....................................................................................46.0mg
L -Histidine·HCl·H 2 O................................................................42.0mg
L -Phenylalanine........................................................................32.0mg
L -Cysteine·2HCl.......................................................................31.0mg
L -Methionine............................................................................15.0mg
L -Glutamic acid........................................................................14.7mg
L -Aspartic acid.........................................................................13.3mg
L -Asparagine·H 2 O....................................................................13.2mg
L -Proline...................................................................................11.5mg
L -Serine....................................................................................10.5mg
L -Tryptophan............................................................................10.0mg
L -Alanine....................................................................................8.9mg
Glycine.......................................................................................7.5mg
Preparation of Amino Acid Solution : Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Filter sterilize.
Vitamin Solution:
Composition per 100.0mL:
i-Inositol.....................................................................................2.0mg
D -Ca pantothenate......................................................................1.0mg
Choline chloride.........................................................................1.0mg
Folic acid....................................................................................1.0mg
Niacinamide...............................................................................1.0mg
Pyridoxal·HCl............................................................................1.0mg
Thiamine·HCl ............................................................................1.0mg
Riboflavin ..................................................................................0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Minimum Essential Medium (MEM): Asepti-
cally combine 400.0mL of sterile inorganic salt solution, 400.0mL of
sterile other component solution, 100.0mL of sterile amino acid solu-
tion, and 100.0mL of sterile vitamin solution.
NaHCO 3 Solution:
Composition per 40.0mL:
NaHCO 3 ........................................................................................2.0g
Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de-
ionized water and bring volume to 40.0mL. Mix thoroughly. Filter ster-
ilize.
Penicillin-Streptomycin Solution
Composition per 10.0mL:
Penicillin.....................................................................................0.01g
Streptomycin...............................................................................0.01g
Preparation of Penicillin-Streptomycin Solution: Add compo-
nents to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Filter sterilize.
Preparation of Medium: Aseptically combine 950.0mL of sterile
minimum essential medium, 100.0mL of sterile heat inactivated fetal
bovine serum, 40.0mL of sterile NaHCO 3 solution, and 10.0mL of ster-
ile penicillin-streptomycin solution. Adjust pH to 7.2 with humidified
10% CO 2 in 90% air.
Use: For the cultivation of Encephalitozoon cuniculi, Encephalito-
zoon hellem, Encephalitozoon intestinalis, Naegleria fowleri, and
Nosema corneum.