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微信小章| 产品名称： | Prototheca zopfii Kruger |
|---|---|
| 商品货号： | TS162281 |
| Strain Designations： | 62-344 UTEX 1442 |
| 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： | digested sludge, Dayton, OH, 1962 |
| Product Format： | freeze-dried |
| Type Strain： | no |
| Medium： | ATCC® Medium 28: Emmons modification of Sabourauds agar |
| Growth Conditions： | Temperature: 25.0°C Duration: axenic Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface. |
| Subcultivation： | Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface. |
| Cryopreservation： | 1.xa0xa0 Harvest cells from a culture which is at or near peak density by adding 3.0-5.0 ml fresh ATCC medium 28 broth to the slant or plate and washing cells into suspension.xa0 It may be helpful to rub the surface of the agar with a spread bar or inoculating loop to detach adhering cells. 2.xa0xa0 Adjust the concentration of cells to 2 x 107/ml with fresh broth medium, then dilute to half this concentration by adding an equal amount of a 20% (v/v) sterile solution of either DMSO or glycerol in fresh ATCC medium 28 broth. 3.xa0xa0 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 preparation and the cryoprotective solution to the start of the cooling cycle should be no less than 15 min and no greater than 30 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 until thawed (2-3 min). Immerse the ampule enough to cover only the frozen material.xa0 Do not agitate the ampule. 7.xa0xa0 Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and add to a fresh slant of ATCC medium 28 or the surface of an agar plate of ATCC medium 28. 8.xa0xa0 Maintain as described above. |
| Name of Depositor： | WB Cooke |
| Year of Origin： | 1962 |
| References： | Pore RS. Prototheca taxonomy. Mycopathologia 90: 129-139, 1985. |