| 产品名称: |
Naegleria morganensis Dobson et al. |
| 商品货号: |
TS165426 |
| Deposited As: |
Naegleria sp. |
| Strain Designations: |
SWL NG-236 |
| Biosafety Level: |
1
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: |
Fresh water, River Murray, Morgan, Australia, 1984 |
| 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: |
yes |
| Comments: |
Zymodeme 44-2 Equals NG-236, maximum temperature tolerance 44C |
| Medium: |
ATCC® Medium 997: Fresh water ameba medium
ATCC® Medium 711: PYB
|
| Growth Conditions: |
Temperature: 25°C |
| Cryopreservation: |
Reagents
Cryoprotective Solution
DMSO, 1.5 mL
Pages Balanced Salt Solution (or similar), 8.5 mL
Harvest and Preservationxa0
- 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 (Pages Balanced Salt Solution) and washing cells into suspension.xa0 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.xa0 Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately -1°C/min.) xa0
- 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.xa0 Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0xa0 Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.xa0 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: |
BS Robinson |
| Year of Origin: |
1984 |
| References: |
Adams M, et al. A genetic approach to species criteria in the amoeba genus Naegleria using allozyme electrophoresis. Int. J. Parasitol. 19: 823-834, 1989. PubMed: 2635158
Dobson PJ, et al. New thermophilic Naegleria species (Heterolobosea: Vahlkampfiidae) from Australia and Asia: allozyme, morphometric and physiological characterization. Acta Protozool. 36: 261-271, 1997.
type strain
De Jonckheere JF. A century of research on the amoeboflagellate genus Naegleria. Acta Protozool. 41: 309-342, 2002.
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