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微信小章| 产品名称： | Anophryoides soldoi Small and Lynn |
|---|---|
| 商品货号： | TS177426 |
| Strain Designations： | 116-1 |
| 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： | coastal waters, Naples, FL, 1971 |
| Product Format： | frozen |
| Type Strain： | no |
| Comments： | Prey-induced transformation |
| Medium： | ATCC® Medium 1651: MA medium |
| Growth Conditions： | Temperature: 25.0°C Duration: axenic Protocol: ATCCNO: 50204 SPEC: Aseptically transfer 0.1 ml of the culture to 5.0 ml of fresh medium in a 16 x 125 mm screw-capped test tube. Incubate cultures upright at 25C with the caps on loosely. Transfer cultures every two weeks. |
| Subcultivation： | Protocol: ATCCNO: 50204 SPEC: Aseptically transfer 0.1 ml of the culture to 5.0 ml of fresh medium in a 16 x 125 mm screw-capped test tube. Incubate cultures upright at 25C with the caps on loosely. Transfer cultures every two weeks. |
| Cryopreservation： | 1.xa0 Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min. 2.xa0 Adjust the concentration of cells to 2 x 106 - 2 x 107/ml in fresh medium. 3.xa0 While cells are centrifuging prepare a 22% (v/v) solution of sterile DMSO in fresh medium. 4. Mix the cell preparation and the 22% DMSO in equal portions. Thus, the final concentration will be 106 - 107 cells/ml and 11% (v/v) DMSO. The time from the 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. 5.xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6.xa0xa0 Place the vials in a controlled rate freezing unit.xa0 From room temperature cool at -1°C/min to -40°C.xa0 If the freezing unit can compensate for the heat of fusion, maintain rate atxa0xa0xa0xa0xa0xa0xa0 -1°C/min through the heat of fusion.xa0 At -40°C plunge 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 xa0xa0xa0xa0xa0 -1°C/min.) xa0 7.xa0 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 should not be stored above -55°C. 8.xa0xa0 To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.xa0 Do not agitate the ampule.xa0 Do not leave ampule in water bath after thawed. 9.xa0xa0 Immediately after thawing, aseptically transfer contents to a 16 x 125 mm screw-capped test tube containing 5 ml of ATCC Medium 1651. 10.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Incubate the tube on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 25°C. |
| Name of Depositor： | AT Soldo, EB Small |
| Year of Origin： | 1971 |
| References： | Gomez-Saladin E, Small E. Prey induced transformation of Miamiensis avidus strain Ma/2 by a soluble factor. J. Eukaryot. Microbiol. 40: 550-556, 1993. |