MPHM Medium

[所属分类:培养基配方] [发布时间:2021-6-8] [发布人:网站管理员2] [阅读次数:] [返回]

MPHM Medium

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

Composition per 1275.0mL:

McCoy's 5A medium, modified....................................................1.0L
Peptone solution.....................................................................250.0mL
Hemin solution.........................................................................25.0mL
Source: McCoy’s 5A medium, modified, is available from Gibco.
McCoys 5A Medium, Modified:
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............................................................................................6.46g
NaHCO 3 ........................................................................................2.2g
NaH 2 PO 4 ·H 2 O.............................................................................0.58g
KCl................................................................................................0.4g
CaCl 2 , anhydrous..........................................................................0.1g
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......................................................................................3.0g
Peptone .........................................................................................0.6g
Phenol Red....................................................................................0.1g
Glutathione, reduced..................................................................0.5mg
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...........................................................................219.2mg
L -Asparagine............................................................................45.0mg
L -Arginine·HCl ........................................................................42.1mg
L -Isoleucine..............................................................................39.3mg
L -Leucine.................................................................................39.3mg
L -Lysine·HCl............................................................................36.5mg
L -Cysteine................................................................................31.5mg
L -Serine....................................................................................26.3mg
L -Tyrosine·2Na·2H 2 O ..............................................................26.2mg
L -Glutamic acid........................................................................22.1mg
L -Histidine·HCl·H 2 O................................................................21.0mg
L -Aspartic acid.........................................................................20.0mg
L -Hydroxyproline.....................................................................19.7mg
L -Threonine..............................................................................17.9mg
L -Valine....................................................................................17.6mg
L -Proline...................................................................................17.3mg
L -Phenylalanine........................................................................16.5mg
L -Methionine............................................................................15.0mg
L -Alanine..................................................................................13.9mg
Glycine.......................................................................................7.5mg
L -Tryptophan..............................................................................3.1mg
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...................................................................................36.0mg
Folic acid..................................................................................10.0mg
Choline chloride.........................................................................5.0mg
Vitamin B 12 ................................................................................2.0mg
p-Aminobenzoic acid.................................................................1.0mg
Ascorbic acid .............................................................................0.5mg
Niacinamide...............................................................................0.5mg
Nicotinic acid.............................................................................0.5mg
Pyridoxal·HCl............................................................................0.5mg
Pyridoxine·HCl..........................................................................0.5mg
Biotin .........................................................................................0.2mg
D -Ca pantothenate......................................................................0.2mg
Riboflavin ..................................................................................0.2mg
Thiamine·HCl ............................................................................0.2mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of McCoys 5A Medium, Modified: Aseptically
combine 400.0mL of sterile inorganic salt solution, 400.0mL of sterile
other component solution, 100.0mL of sterile amino acid solution, and
100.0mL of sterile vitamin solution.
Peptone Solution:
Composition per 300.0mL:
Proteose peptone No. 3.................................................................7.5g
Preparation of Peptone Solution: Add components to distilled/
deionized water and bring volume to 300.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Hemin Solution:
Composition per 100.0mL:
NaCl..............................................................................................0.8g
Na 2 HPO 4 .....................................................................................0.12g
Hemin .........................................................................................0.05g
KCl..............................................................................................0.04g
MgCl 2 ·6H 2 O ...............................................................................0.03g
NaH 2 PO 4 ·H 2 O.............................................................................0.02g
CaCl 2 ......................................................................................... 0.011g
Preparation of Hemin Solution: Add NaCl, KCl, NaH 2 PO 4 ·H 2 O,
and Na 2 HPO 4  to distilled/deionized water and bring volume to
50.0mL. Mix thoroughly. Separately add the CaCl 2 and MgCl 2 ·6H 2 O
to distilled/deionized water and bring volume to 50.0mL. Mix thor-
oughly. Combine the two solutions. Mix thoroughly. Add 0.05g of
hemin. Mix thoroughly. Adjust to pH 9.0 with 10N NaOH to dissolve
the hemin. Mix thoroughly. Filter sterilize.
Preparation of Medium: Aseptically combine 1.0L of sterile mod-
ified McCoy’s medium 5A, 250.0mL of sterile peptone solution, and
25.0mL of sterile hemin solution. Aseptically distribute into sterile
screw-capped tubes or flasks.
Use:  For the cultivation of Trypanosoma theileri.