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
Division mutant derived from wild type strain UTEX 1232, 1987
Product Format
frozen
Storage Conditions
Frozen: -70°C or colder Freeze-Dried: 2°C to 8°C Live Culture: See Protocols Section
Type Strain
no
Disclosure
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC.
Comments
Carotenoid producing culture
Medium
ATCC® Medium 2001: CYU 2%
Growth Conditions
Temperature: 25°C to35°C
Cryopreservation
Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.
Adjust the concentration of cells to 2 x 106 - 2 x 107/mL in fresh medium.
While cells are centrifuging prepare a solution of 14% (v/v) sterile DMSO solution in fresh medium.
Mix the cell preparation and the cryoprotective solution in equal portions. Thus, the final concentration will be 106 - 107 cells/mL and 7% (v/v) DMSO. The time from the mixing of the cell preparation and cryoprotective stock solution to the start of the cooling cycle should be no less than 5 min and no greater than 15 min.
Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
Place the vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion. At -40°C plunge 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.)
The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials should not be stored above -55°C.
To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial just to a level just above the surface of the frozen material. Do not agitate the vial.
Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and add to a centrifuge tube containing 5 mL of ATCC medium 2001.
Incubate the culture at 50-100 µEinsteins/m2/s irradiance at 25-35°C. Maintain under a 14/10h light-dark photoperiod.
