PDM-114 Medium

[所属分类:培养基配方] [发布时间:2021-6-30] [发布人:网站管理员2] [阅读次数:] [返回]

PDM-114 Medium

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

Composition per liter:

Base solution..........................................................................770.0mL
Bovine serum, heat inactivated..............................................100.0mL
Solution A................................................................................50.0mL
Solution B................................................................................50.0mL
Special 107 vitamin mix..........................................................30.0mL
pH 6.8 ± 0.2 at 25°C
Base Solution:
Composition per 770.0mL:
Pancreatic digest of casein..........................................................20.0g
NaCl............................................................................................. 2.0g
L -Cysteine·HCl..............................................................................1.0g
Ascorbic acid ............................................................................... 0.2g
Ammonium chloride...................................................................0.16g
Adenine....................................................................................73.0mg
Guanosine ................................................................................59.0mg
Adenosine 5′-monophosphate .................................................52.0mg
Uracil .......................................................................................37.0mg
Cytosine.................................................................................. 30.0mg
Ferric ammonium citrate..........................................................22.8mg
Adenosine 5′-diphosphate .......................................................16.6mg
Adenosine 5′-triphosphate.........................................................7.6mg
Preparation of Base Solution: Add components to distilled/deion-
ized water and bring volume to 770.0mL. Mix thoroughly. Adjust pH
to 6.8 with NaOH. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Solution A:
Composition per 50.0.0mL:
Glucose .......................................................................................10.0g
Preparation of Solution A: Add glucose to distilled/deionized wa-
ter and bring volume to 50.0mL. Mix thoroughly. Filter sterilize.
Solution B:
Composition per 50.0.0mL:
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................0.6g
Preparation of Solution B: Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature.
Special 107 Vitamin Mix:
Composition per 120.0.0mL:
Solution 4 vitamins................................................................100.0mL
Solution 2...................................................................................1.2mL
Solution 1...................................................................................0.4mL
Solution 3...................................................................................0.4mL
Solution 1:
Composition per 100.0mL:
Absolute ethanol....................................................................100.0mL
DL -6,8-Thioctic acid, oxidized...............................................100.0mg
Preparation of Solution 1: Add  DL -6,8-thioctic acid to absolute
ethanol and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Solution 2:
Composition per 100.0mL:
Vitamin B 12 ..............................................................................40.0mg
Preparation of Solution 2: Add vitamin B 12 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Solution 3:
Tween™ 80.................................................................................50.0g
Absolute ethanol....................................................................100.0mL
Preparation of Solution 3: Add Tween™ 80 to absolute ethanol
and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Solution 4 Vitamins:
Composition per liter:
Vitamin B 12 ..............................................................................10.0mg
Calcium  D -(+)-pantothenate ......................................................2.3mg
Choline chloride.......................................................................1.25mg
Riboflavin..................................................................................0.7mg
Vitamin A, crystallized alcohol ...............................................0.25mg
Calciferol (vitamin D 2 )............................................................0.25mg
i-Inositol.................................................................................0.125mg
p-Aminobenzoic acid.............................................................0.125mg
Niacin...................................................................................0.0625mg
Niacinamide.........................................................................0.0625mg
Pyridoxal·HCl......................................................................0.0625mg
Pyridoxine·HCl....................................................................0.0625mg
α-Tocopherol phosphate, disodium salt.................................0.025mg
Biotin.....................................................................................0.025mg
Folic acid ...............................................................................0.025mg
Menadione (vitamin K 3 )........................................................0.025mg
Thiamine·HCl........................................................................0.025mg
Preparation of Solution 4 Vitamins: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Preparation of Special 107 Vitamin Mix: Aseptically combine
0.4mL of sterile solution 1, 1.2mL of sterile solution 2, 0.4mL of sterile
solution 3, and 100.0mL of sterile solution 4 vitamins. Bring volume
to 120.0mL with sterile distilled/deionized water.
Preparation of Medium:  Aseptically combine 770.0mL of sterile
base solution, 100.0mL of heat-inactivated bovine serum, 50.0mL of
sterile solution A, 50.0mL of sterile solution B, and 30.0mL of sterile
special 107 vitamin mix. Aseptically distribute into sterile tubes or
flasks.
Use: For the cultivation of Entamoeba histolytica.