| 產(chǎn)品名稱(chēng) |
Naegleria fowleri Carter |
| 商品貨號(hào) |
B241036 |
| Deposited As |
Echinostelium sp. |
| Strain Designations |
B-L |
| Biosafety Level |
2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Isolation |
Cerebrospinal fluid of 7-year-old boy, Matamath, New Zealand |
| Product Format |
frozen |
| Storage Conditions |
Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures:
2-8°C
Live Cultures:
See Protocols section for handling information |
| Type Strain |
no |
| Comments |
Biochemical identification |
| Medium |
ATCC® Medium 997: Fresh water ameba medium
ATCC® Medium 711: PYB
ATCC® Medium 1323: Page's balanced salt solution (PBS)
|
| Growth Conditions |
Temperature: 25°C
Culture System: Xenic |
| Cryopreservation |
Reagents
Cryoprotective Solution
DMSO 1.5 mL
Page's Balanced Salt Solution (or similar) 8.5 mL
Harvest and Preservation
- Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.
- Harvest cells from a culture which is at or near peak density by adding 5 mL ATCC medium 1323 (Page's Balanced Salt Solution) and washing cells into suspension. Rub the surface of the plate with a spread bar to detach adhering trophozoites.
- Adjust the concentration of cells to at least 2 x 106/mL in fresh medium.
- Mix the cell preparation and the cryoprotective solution in equal portions.
- Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
- Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)
- Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
- To establish a culture from the frozen state place the vial in a 35°C water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997. Distribute the material evenly over the plate using a spread bar.
- Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.
- Follow the protocol for maintenance of culture.
|
| Name of Depositor |
BN Mandal |
| References |
Daggett PM, Nerad TA. The biochemical identification of vahlkampfiid amoebae. J. Protozool. 30: 126-128, 1983. PubMed: 6864593
N.Z. Med. J. 71: 16-23, 1970.
De Jonckheere JF. A century of research on the amoeboflagellate genus Naegleria. Acta Protozool. 41: 309-342, 2002.
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