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
Seawater, 1976
Product Format
test tube
Storage Conditions
Frozen Cultures: -70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures: 2-8°C
Live Cultures: See Protocols section for handling information
Type Strain
no
Comments
Photosynthetic. Nonaxenic.
Medium
ATCC® Medium 1405: HESNW medium
Growth Conditions
Temperature: 20°C to 25°C
Atmosphere: Aerobic
Cryopreservation
Reagents
Cryoprotective Solution
Glycerol 2.4 mL
Fresh growth medium w/o bacteria 7.6 mL
Harvest and Preservation
Mix the components of the cryoprotective solution in the order listed.
Harvest cells from a culture that is at or near peak density by centrifugation at 800-1000 x g for 5 min.
Adjust the concentration of cells to at least 2 x 107/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 16 x 125 mm screw-capped test tube containing 5 mL of sterile ATCC medium 1405. Immediately seal and agitate the culture to evenly suspend cells, then aseptically transfer a 0.5 mL aliquot to a second, identical tube of medium.
Incubate the parent and daughter cultures at a 15° horizontal slant at 20-25°C with the caps on loosely for air exchange. Maintain under a 14 hour light (~50 µEinsteins/m2/s irradiance)/10 hour dark cycle.
Follow the protocol for maintenance of culture.
Name of Depositor
University of British Columbia
Chain of Custody
ATCC <-- University of British Columbia <-- R. Waters
