Perkinsus Agar Medium (ATCC Medium 2289)

[所属分类:培养基配方] [发布时间:2021-7-5] [发布人:网站管理员2] [阅读次数:] [返回]

Perkinsus Agar Medium
(ATCC Medium 2289)

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

Composition per liter:

Modified Perkinsus Medium .................................................485.0mL
Agar Medium.........................................................................485.0mL
Fetal bovine serum, heat inactivated .......................................20.0mL
Lipid concentrate.....................................................................10.0mL
Modified Perkinsus Medium
Composition per liter:
HEPES...................................................................................... 11.92g
Nutrient mix F-12 Ham ..............................................................10.8g
Dulbecco’s modified Eagle’s medium..........................................8.4g
L -Glutamine................................................................................0.29g
NaHCO 3 ........................................................................................1.3g
SASW 2X solution ................................................................860.0mL
JPL carbohydrate solution .......................................................20.0mL
Phenol Red (0.5%) solution.......................................................1.0mL
SASW 2X Solution:
Composition per liter:
Seawater, synthetic basal mixture...............................................36.4g
Preparation of 2X SASW Solution: Add seawater synthetic basal
mixture to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly.
JLP Carbohydrate Solution:
Composition per 100.0mL:
Glucose.........................................................................................5.0g
Galactose.......................................................................................1.0g
Trehalose.......................................................................................1.0g
Preparation of JLP Carbohydrate Solution: Add components
to distilled/deionized water and bring volume to 100.0mL. Mix thor-
oughly.
Phenol Red Solution:
Composition per 10.0mL:
Phenol Red..................................................................................0.05g
Preparation of Phenol Red Solution:  Add Phenol Red to dis-
tilled/deionized water and bring volume to 10.0mL.
Preparation of Modified Perkinsus Medium:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize
Agar Medium:
Composition per liter:
Agar............................................................................................30.0g
Preparation of  Agar  Medium: Add agar to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Gently heat while stir-
ring and bring to boiling. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 45°–50°C.
Lipid Concentrate:
Composition per liter:
Pluronic™ F68............................................................................10.0g
Tween™ 80...................................................................................2.5g
Cod liver oil..................................................................................1.0g
Cholesterol..................................................................................0.45g
DL -α-Tocopherol acetate...............................................................0.2g
Preparation of Lipid Concentrate: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Warm Modified Perkinsus Medium to
50°C and combine with Agar Medium at 50°C. Mix thoroughly and
maintain at 50°C. Aseptically add heat-inactivated fetal bovine serum
and lipid concentrate. Mix thoroughly. Aliquot in 20mL amounts to Pe-
tri dishes and allow to solidify.
Use: For the cultivation of Perkinsus marinus, P. andrewsi, P. chesa-
peaki, and P. atlanticus.