Tetraselmis sp. (ATCC® 50246)

Strain Designations: PB25  /  Depositor: N Yubuki  /  Biosafety Level: 1

Permits and Restrictions

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Strain Designations PB25
Application
Biofuel production
Biosafety Level 1
Isolation Tide pool, Pachena Beach, British Columbia, Canada, 2010
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Comments
Food source for Rapaza viridis
Medium Medium 1747: Tetraselmis medium
Growth Conditions
Temperature: 25°C
Culture System: Axenic
Cryopreservation Reagents
Cryoprotective Solution
DMSO, 1.5 mL
Fresh growth medium, 8.5 mL

Harvest and Preservation
  1. Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.
  2. Adjust the concentration of cells to 2 x 106 - 2 x 107/mL in fresh medium.
  3. While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO in fresh medium.
  4. Mix the cell preparation and the 15% DMSO in equal portions. Thus, the final concentration will be 106 - 107 cells/mL and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution to the beginning of the freezing process should be no less than 5 min and no greater than 15 min.
  5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  6. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion.  At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  
  7. 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 should not be stored above -55°C.
  8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial just to a level just above the surface of the frozen material. Do not agitate the vial.
  9. Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and add to a centrifuge tube containing 5 mL of ATCC medium 1747.  Centrifuge at 300 x g for 5 min.
  10. Remove most of the supernatant (=cryoprotectant, which can inhibit growth) and then resuspend the pellet.  Transfer the culture to a 16 x 125 mm screw-capped test tube containing 5 mL of ATCC medium 1747.
  11. Incubate on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 25°C under a 14 hour light (~50 µEinsteins/m2/s irradiance)/10 hour dark cycle.
Name of Depositor N Yubuki
Year of Origin 2010
References

Yamaguchi A, et al. Morphostasis in a novel eukaryote illuminates the evolutionary transition from phagotrophy to phototrophy: description of Rapaza viridis n. gen. et sp. (Euglenozoa, Euglenida). BMC Evol. Biol. 8: 12-29, 2012. PubMed: 22401606

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation