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微信小章| 产品名称： | Crypthecodinium cohnii (Seligo) Javornicky |
|---|---|
| 商品货号： | TS142387 |
| Strain Designations： | Da1 |
| Application： | Biofuel production |
| 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： | Sargassum sp., Daytona Beach, FL |
| Product Format： | frozen |
| Type Strain： | no |
| Comments： | Minor Sibling Species Da |
| Medium： | ATCC® Medium 460: A2E6 medium |
| Growth Conditions： | Temperature: 25.0°C Duration: axenic Protocol: ATCCNO: 30021 SPEC: This culture is routinely shipped as a frozen stabilate. Thaw the ampule and aseptically transfer the material to 5 ml of ATCC medium 460 in a 16 x 125 mm screw-capped test tube. Do not distribute the thawed material to a larger volume of medium. It is essential to first establish the culture in a small volume. Screw cap on tightly, loosen one half turn, and incubate culture upright at 25C. The culture should be ready to subculture in approximately 7 days. To subculture, screw the cap on tightly, invert the culture 5 times and aseptically transfer a 0.1 ml aliquot to 5 ml of ATCC medium 460 and incubate as above. Prepare two subcultures weekly. Retain all cultures for up to one month to ensure against loss. |
| Subcultivation： | Protocol: ATCCNO: 30021 SPEC: This culture is routinely shipped as a frozen stabilate. Thaw the ampule and aseptically transfer the material to 5 ml of ATCC medium 460 in a 16 x 125 mm screw-capped test tube. Do not distribute the thawed material to a larger volume of medium. It is essential to first establish the culture in a small volume. Screw cap on tightly, loosen one half turn, and incubate culture upright at 25C. The culture should be ready to subculture in approximately 7 days. To subculture, screw the cap on tightly, invert the culture 5 times and aseptically transfer a 0.1 ml aliquot to 5 ml of ATCC medium 460 and incubate as above. Prepare two subcultures weekly. Retain all cultures for up to one month to ensure against loss. |
| Cryopreservation： | 1.xa0xa0 Harvest cells from cultures which are at or near peak density.xa0 Aseptically transfer cells to 15 ml plastic centrifuge tubes and centrifuge at ~150 x g for 5 min. 2. Adjust the concentration of cells to 2 x 106/ml with fresh medium, then dilute to half this concentration by adding an equal amount of a 15% (v/v) sterile glycerol solution in freshxa0 medium. 3.xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).xa0 The time from mixing of the cell suspension and the glycerol solution, before the cooling cycle begins, should be no greater than 15 min. 4.xa0xa0 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.) xa0xa0 5.xa0xa0 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. 6.xa0xa0 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. 7.xa0xa0 Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and transfer to a fresh tube containing 5 ml of ATCC Medium 460. Incubate the tube vertically at 20-25°C with the cap loosened one half turn. xa0Subculture every 10-14d. |
| Name of Depositor： | CA Beam, M Himes |
| References： | Beam CA, Himes M. Electrophoretic characterization of members of the Crypthecodinium cohnii (Dinophyceae) species complex. J. Protozool. 34: 204-217, 1987. |