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微信小章| 产品名称： | Naegleria fowleri Carter |
|---|---|
| 商品货号： | TS142189 |
| Strain Designations： | PA-90 |
| Biosafety Level： | 2
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： | domestic water supply, Port Augusta, Australia, 1972 |
| Product Format： | frozen |
| Type Strain： | no |
| Comments： | use of plastic ampoules for freeze preservation |
| Medium： | ATCC® Medium 1034: Modified PYNFH medium (Available from ATCC as ATCC cat. no. 327-X) |
| Growth Conditions： | Temperature: 35.0°C Duration: axenic |
| Cryopreservation： | 1.xa0 Harvest cells from a culture that is at or near peak density by centrifugation at 600 x g for 5 min. Pool the cell pellets into a single tube. 2.xa0 Adjust the concentration of cells to 2.0 x 106/ml.xa0 If the concentration is too low, centrifuge at 600 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired concentration. 3.xa0xa0 Prepare a 15% (v/v) sterile DMSO solution in ATCC medium 1034 as follows:xa0 Add the required volume of DMSO to a glass screw-capped test tube and place on ice.xa0 Allow the DMSO to solidify.xa0 Add the required volume of refrigerated ATCC medium 1034.xa0 Dissolve the DMSO by inverting several times.xa0 If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium. 4.xa0 Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 106 and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 60 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.xa0 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 the ampules into liquid nitrogen. 7.xa0 The frozen preparations are stored in either the vapor or liquid phase of a nitrogen refrigerator. 8.xa0xa0 To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial enough to cover only the frozen material. Do not agitate the vial. 9.xa0xa0 Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate into 5.0 ml of fresh ATCC medium 1034. 10.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Incubate the tube on a 15° horizontal at 35°C with the cap screwed on tightly. |
| Name of Depositor： | JA Jamieson |
| Year of Origin： | 1972 |
| References： | Anderson K, Jamieson A. Agglutination test for the investigation of the genus Naegleria. Pathology 4: 273-278, 1972. PubMed: 4641057 Simione FP Jr., et al. The use of plastic ampoules for freeze preservation of microorganisms. Cryobiology 14: 500-502, 1977. PubMed: 891238 |