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
Sediment from man-made marsh island at edge of dredge spoilage site, Beaufort, NC
Product Format
frozen
Storage Conditions
Frozen: -70°C or colder Freeze-Dried: 2°C to 8°C Live Culture: See Protocols Section
Type Strain
yes
Comments
Originally accessioned as Corallomyxa sp.
Taxonomy
Medium
ATCC® Medium 1525: Seawater 802 medium
Growth Conditions
Temperature: 25°C
Culture System: Xenic, grown with Klebsiella pneumoniae subsp. pneumoniaeATCC 700831 or Enterobacter aerogenes ATCC 13048
Cryopreservation
Reagents
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium w/o bacteria, 8.0 mL
Harvest and Preservation
Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.
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 to at least 2 x 106/mL in fresh 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. 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 at 25°C with the cap screwed on tightly.
Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 mL to 10.0 mL of bacterized ATCC medium 1525.
Follow the protocol for maintenance of culture.
Name of Depositor
TA Nerad
References
Bass D, et al. Phylogeny of novel naked filose and reticulose Cercozoa: Granofilosea cl. n. and Proteomyxidea revised. Protist 160: 75-109, 2009. PubMed: 18952499
Thomas A Nerad, personal communication
Grant JR, et al. Gene discovery from a pilot study of the transcriptomes from three diverse microbial eukaryotes: Corallomyxa tenera, Chilodonella uncinata, and Subulatomonas tetraspora. Protist Genomics 1: 3-18, 2012.
Cross References
Nucleotide (GenBank) :
EF514503
18S small subunit ribosomal RNA gene
