Diaphanoeca grandis Ellis (ATCC® 50111)

Organism: Diaphanoeca grandis Ellis  /  Depositor: PG Davis

Permits and Restrictions

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Strain Designations 1/19/82 NB (3)
Application
Phylogenetic analysis of SSU rRNA, LSU rRNA, tubA, and hsp-90
Biosafety Level 1
Isolation
Narragansett Bay, RI, 1982
Product Format test tube
Type Strain no
Comments
phylogeny
Phylogenetic analysis of SSU rRNA, LSU rRNA, tubA, and hsp-90
Medium ATCC® Medium 1361: Marine flagellate medium
Growth Conditions
Max Temperature: 18.0°C
Min Temperature: 4.0°C
Duration: grown with mixed bacteria
Protocol: ATCCNO: 50111 SPEC: Upon receipt, remove test tube from package and immediately place it between 4 and 18C. Do not allow it to reach room temperature. Also place a T-25 tissue culture flask containing 10 ml of fresh medium and 1 or 2 autoclaved rice grains at the same temperature. Allow the vessels to equilibrate for 2-3 hours and then aseptically transfer a 0.1 ml aliquot from the test tube to the flask. Maintain both culture vessels. Transfer every 14-21 days when maintained at 10C.
Subcultivation
Protocol: ATCCNO: 50111 SPEC: Upon receipt, remove test tube from package and immediately place it between 4 and 18C. Do not allow it to reach room temperature. Also place a T-25 tissue culture flask containing 10 ml of fresh medium and 1 or 2 autoclaved rice grains at the same temperature. Allow the vessels to equilibrate for 2-3 hours and then aseptically transfer a 0.1 ml aliquot from the test tube to the flask. Maintain both culture vessels. Transfer every 14-21 days when maintained at 10C.
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.  Cool on ice prior to use.

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 at least 2 x 106/ml in fresh, ice-cold 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.  Do not allow ampule to overheat.  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 ice-cold ATCC medium 1405 and two sterile rice grains.

9.     Incubate at 4-18°C with the cap screwed on tightly.

10.   Once the culture is established, follow the protocol for maintenance of culture.

Name of Depositor PG Davis
Year of Origin 1982
References

Wainright PO, et al. Monophyletic origins of the metazoa: an evolutionary link with fungi. Science 260: 340-342, 1993. PubMed: 8469985

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

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