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微信小章| 产品名称： | Diplonema sp. 5 |
|---|---|
| 商品货号： | TS133063 |
| Strain Designations： | VIIIB |
| 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： | marine water, Cliff Pool, Bermuda, 1986 |
| Product Format： | frozen |
| Type Strain： | no |
| Medium： | ATCC® Medium 1728: Enriched Isonema medium |
| Growth Conditions： | Temperature: 25.0°C Duration: axenic |
| Cryopreservation： | 1.xa0xa0 Harvest the culture by agitating the contents of each flask.xa0 Transfer the cell suspensions to 15 ml plastic centrifuge tubes. 2.xa0xa0 Spin the cell suspensions at approximately 500 x g for 5 min. 3.xa0xa0 Pool the cell pellets and adjust the concentration to 2.0 - 4.0 x 107 cells/ml with a fresh ATCC medium 1728. xa0xa0xa0xa0xa0 *If the concentration is too low centrifuge at 500 x g for 5 min and resuspend in the volume of ATCC medium 1728 required to yield the desired concentration. 4.xa0xa0 Mix the cell preparation and 20% (v/v) DMSO in equal portions.xa0 The final concentration will be 1.0 - 2.0 x 107 cells/ml and 10% DMSO.xa0 The time from the mixing of the cell preparation and cryoprotective solution to the start of the freezing process should be no less than 15 min. and no more than 30 min. 5.xa0xa0 Dispense in 0.5 ml aliquots to 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 2.5 to 3 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately -1°C/min.) xa0 7.xa0xa0 Store in either the vapor or liquid phase of a nitrogen refrigerator. 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 T-25 tissue culture flask containing 10 ml ATCC medium 1728. 10.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Incubate in a 25°C CO2 incubator with the caps screwed on tightly. |
| Name of Depositor： | TA Nerad |
| Year of Origin： | 1986 |