Phylogenetic analysis of SSU rRNA, LSU rRNA, tubA, and hsp-90
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
Narragansett Bay, RI, 1982
Product Format
test tube
Storage Conditions
Test Tube: See handling procedure
Type Strain
no
Comments
phylogeny
Phylogenetic analysis of SSU rRNA, LSU rRNA, tubA, and hsp-90
Medium
ATCC® Medium 1405: HESNW medium
ATCC® Medium 1361: Marine flagellate medium
Growth Conditions
Temperature: 4-18°C (best growth is achieved at 8-10°C)
Cryopreservation
Cryoprotective Solution
DMSO, 2.0 ml
Fresh growth medium w/o bacteria, 8.0 ml
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.
Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.
Adjust the concentration of cells at least 2 x 106/ml in fresh, ice-cold medium.
Mix the cell preparation and the cryoprotective solution in equal portions.
Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
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.)
Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
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.
Incubate at 4-18°C with the cap screwed on tightly.
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
