Stephanoeca diplocostata Ellis (ATCC® PRA-392)

Organism: Stephanoeca diplocostata Ellis  /  Depositor: N King

Permits and Restrictions

View Permits

Strain Designations Australia
Biosafety Level 1
Isolation Seawater sample, Swan Bay, Victoria, Australia, 1996
Type Strain no
Comments This item is presently archived as original material and is available on a "made-to-order" basis.
Sequenced as part of a choanoflagellate transcriptome project.
Medium ATCC® Medium 1361: Marine flagellate medium
Growth Conditions Temperature: 15°C
Culture System: Xenic culture
Cryopreservation Cryoprotective Solution

DMSO,  2.0 mL

Fresh growth medium w/o bacteria,  8.0 mL

 

1. 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. Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.

3. Adjust the concentration of cells to at least 2 x 106/mL in fresh medium.

4. Mix the cell preparation and the cryoprotective solution in equal portions.

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 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.  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. Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8. 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 of uninoculated ATCC medium 1361.

9. Incubate at 15°C with the cap screwed on tightly.

10. Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 mL to 10.0 mL of uninoculated ATCC medium 1361.

11. Follow the protocol for maintenance of culture.

Name of Depositor N King
Chain of Custody ATCC <-- N King <-- B Leadbeater
References

Carr M, et al. Molecular phylogeny of choanoflagellates, the sister group to Metazoa. Proc. Natl. Acad. Sci. USA. 105: 16641-16646, 2008. PubMed: 18922774

Cross References

Nucleotide (GenBank) : EU011926 18S rRNA gene

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