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微信小章| 产品名称： | Didinium nasutum (Muller) Stein |
|---|---|
| 商品货号： | TS133638 |
| 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： | freshwater pond, San Francisco, CA, 1958 |
| Product Format： | test tube |
| Type Strain： | no |
| Comments： | Changes in the cell volume Preservation |
| Medium： | ATCC® Medium 802: Sonneborns Paramecium medium |
| Growth Conditions： | Temperature: 25.0°C Protocol: ATCCNO: 30399 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically, add 1 ml of ATCC medium 802 inoculated 24 hours previously with bacteria (e.g. Enterobacter aerogenes ATCC-13048) to the inner shell vial. Once completely rehydrated, aseptically transfer the material to a T-25 tissue culture flask containing a thriving culture of Paramecium, not provided. Place the flask at at 25C. Excystment should occur within a few days. |
| Subcultivation： | Protocol: ATCCNO: 30399 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically, add 1 ml of ATCC medium 802 inoculated 24 hours previously with bacteria (e.g. Enterobacter aerogenes ATCC-13048) to the inner shell vial. Once completely rehydrated, aseptically transfer the material to a T-25 tissue culture flask containing a thriving culture of Paramecium, not provided. Place the flask at at 25C. Excystment should occur within a few days. |
| Cryopreservation： | Cryoprotective Solution DMSO xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 2.0 ml Fresh growth medium w/o bacteriaxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 8.0 ml 1.xa0xa0xa0xa0 Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat. 2. xa0xa0 Harvest Didinium cysts from a culture that has recently passed peak density by filtration and centrifugation at 800 x g for 5 min. 3.xa0xa0xa0xa0 Adjust the concentration of cysts at least 2 x 104/ml in fresh medium. 4.xa0 xa0xa0 Mix the cell preparation and the cryoprotective solution in equal portions. 5.xa0 xa0xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6. xa0xa0xa0 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 7.xa0 xa0xa0 Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator. 8.xa0 xa0xa0 To establish a culture from the frozen state place the vial in a 35°C water bath. 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 water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 ml ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048). 9.xa0xa0xa0xa0 Aseptically transfer 1-2 ml from a thriving culture of Paramecium to the T-25 flask. Incubate at 25°C with the cap screwed on tightly. 10.xa0xa0 Once the culture is established, follow the protocol for maintenance of culture. |
| Name of Depositor： | W Balamuth |
| Year of Origin： | 1958 |
| References： | Salt GW. Changes in the cell volume of Didinium nasutum during population increase. J. Protozool. 22: 112-115, 1975. McGrath MS, et al. Studies on the preservation of the ciliate Didinium nasutum. Trans. Am. Microsc. Soc. 96: 519-525, 1977. |