| 產(chǎn)品名稱 |
Helicosporidium sp. |
| 商品貨號 |
B174176 |
| Strain Designations |
Sj-1 |
| 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 |
black fly larvae, Simulium jonesi, Hatchet Creek, Alachua County, FL, 1998 |
| Product Format |
frozen |
| Storage Conditions |
Frozen: -70°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage |
| Axenic/Xenic |
Axenic |
| Type Strain |
no |
| Comments |
pathogenic to black flies development |
| Medium |
ATCC® Medium 28: Emmons' modification of Sabouraud's agar
|
| Growth Conditions |
Temperature: 25°C |
| Cryopreservation |
- Harvest cells from a culture that is at or near peak density. Add 2-3 ml fresh ATCC medium 28 broth to each plate and wash cells into suspension.
- Collect cells by centrifugation at 800 x g for 5 min. Adjust the concentration of cells to 2 x 106 - 2 x 107/ml in fresh medium.
- While cells are centrifuging prepare a 10% (v/v) solution of sterile methanol in fresh broth medium.
- Mix the cell preparation and the 10% methanol solution in equal portions. Thus, the final concentration will be 106 - 107 cells/ml and 5% (v/v) Methanol. The time from mixing of the cell preparation and methanol stock solution to the beginning of the freezing process should be no less than 5 min and no greater than 15 min.
- Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
- Place the vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion. At -40°C plunge 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.)
- The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80 and -70°C for no longer than one week.
- To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.
- Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and transfer to a test tube containing 5 ml of ATCC medium 28 broth or to the surface of an ATCC medium 28 agar plate.
- Incubate a test tube culture upright at 25°C with the cap screwed on loosely (loosened one-half turn); incubate a plate upright at 25°C. Subculture every 4-6 weeks when incubated at 25°C, or every 6-12 months when incubated at 18°C.
|
| Name of Depositor |
DG Boucias |
| Chain of Custody |
ATCC <-- DG Boucias <-- S.E. White |
| Year of Origin |
1998 |
| References |
Boucias DG, et al. In vivo and in vitro development of the protist Helicosporidium sp.. J. Eukaryot. Microbiol. 48: 460-470, 2000. PubMed: 11456323
|
