Blastocystis Egg Medium

[所属分类:培养基配方] [发布时间:2019-12-24] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Blastocystis Egg Medium

培养基配方
Composition per 1300.0mL:
Homogenized whole egg....................................783.0mL
Stone's modification
of Locke's solution .....................................217.0mL
Horse serum, heat inactivated............................300.0mL
Homogenized Whole Egg:
Composition per liter:
Whole eggs ...............................................18–24
Preparation of Homogenized Whole Egg:  Use fresh fertile eggs,
less than 1 week old. Scrub the shells with soap. Let stand in a soap solu-
tion for 30 min. Rinse in running water. Soak eggs in 70% ethanol for 15
min. Break the eggs into a sterile container. Homogenize by shaking. Filter
through four layers of sterile cheesecloth into a sterile graduated cylinder.
Measure out 1.0L.
Stone's Modification of Locke's Solution:
Composition per liter:
NaCl............................................................8.0g
Na 2 HPO 4 .....................................................2.0g
NaHCO 3 ........................................................0.4g
KH 2 PO 4 ......................................................0.3g
CaCl 2 .........................................................0.2g
KCl.............................................................0.2g
MgCl 2 ·6H 2 O ...............................................0.01g
Preparation of Stone's Modification of Locke's Solution:
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly.
Preparation of Medium: Distribute homogenized whole egg in
4.0mL volumes into 16 × 125mm screw-capped test tubes. Place tubes
in a slanted position. Inspissate at 80°C (moist heat) for 10 min. Allow
to cool. Add 4.5mL of Stone's modification of Locke's solution to the
surface of the solidified egg in each tube. Close tubes with a rubber
stopper. Place tubes in a press. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature. Aseptically replace rubber stoppers
with sterile screw caps. Prior to use, aseptically add 1.5mL of heat-in-
activated sterile horse serum to each tube.
Use: For the cultivation of Anophryoides species, Blastocystis hominis,
other Blastocystis species, Endolimax nana, and Metanophrys species.