| 产品名称： |
Hartaetosiga balthica (Wylezich and Karpov) Carr, Richter and Nitsche |
| 商品货号： |
TS175811 |
| Deposited As： |
Monosiga gracilis Kent |
| Strain Designations： |
BSA-02190019 |
| 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： |
Salt marsh sediment, Cattleshed Marsh, eastern shore of Virginia, 2000 |
| Product Format： |
frozen |
| Storage Conditions： |
Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures:
2-8°C
Live Cultures:
See Protocols section for handling information |
| Type Strain： |
no |
| Medium： |
ATCC® Medium 1525: Seawater 802 medium
|
| Growth Conditions： |
Temperature: 25°C
Atmosphere: Aerobic
Culture System: Xenic |
| Cryopreservation： |
Reagents
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium, 8.0 mL
Harvest and Preservation
- To achieve the best results, set up cultures with several different inocula (i.e., 0.5 mL, 1.0 mL, and 2.0mL).xa0 Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density. xa0Use a sterile cotton swab, cell scraper, or a rubber policeman to detach adherent organisms
- Adjust the concentration to approximately 1 x 107 cells/mL by centrifugation at 700-800 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.xa0
- While cells are centrifuging prepare a 20% (v/v) solution of sterile DMSO as follows:xa0 Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.xa0 Allow the DMSO to solidify.xa0 Add the required volume of refrigerated medium.xa0 Dissolve the DMSO by inverting the tube several times.
Note: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
- Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be approximately 5 x 106 cells/mL and 10% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 30 min.
- Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
- 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 at -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 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
- The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.
- To establish a culture from the frozen state, place the vial in a 35°C water bath.xa0 Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0xa0 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 1525 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).
- Incubate with the cap tightly sealed at 20-25°C.
- Once the culture is established, follow the protocol for maintenance of culture.xa0
|
| Name of Depositor： |
TA Nerad |
| Year of Origin： |
2000 |
| References： |
Thomas A Nerad, personal communication
|
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