Ankistrodesmus braunii Brunnthaler (ATCC® 12744)

Organism: Ankistrodesmus braunii Brunnthaler  /  Depositor: RW Krauss

Permits and Restrictions

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Strain Designations P202/78
Application
This strain is recommended by ATCC for use in the tests described in ASTM Standard Test Method E1366-91 where only the taxon is specified.
Biosafety Level 1
Product Format frozen
Type Strain no
Comments
This strain is recommended by ATCC for use in the tests described in ASTM Standard Test Method E1366-91 where only the taxon is specified.
cryopreservation
effect of apatites on growth
photosynthetic
Medium ATCC® Medium 5: Sporulation agar
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Cryopreservation

1.  Harvest cells from a culture that is at or near peak density by centrifugation at 700-800 x g for 5 min.

2.   Adjust the concentration of cells to 2 x 106 - 2 x 107/ml in fresh medium.

3.  While cells are centrifuging prepare a 14% (v/v) solution of sterile DMSO in fresh medium.

4.   Mix the cell preparation and the 14% DMSO 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 DMSO stock solution to the beginning of the freezing process should be no less than 5 min and no greater than 15 min.

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 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.)  

7.   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.

8.   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.

9.   Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and add to a 16 x 125 mm screw-capped test tube containing 5 ml of ATCC medium 5 broth or to the surface of an ATCC medium 5 agar plate (20 x 100 mm Petri plate containing 20 ml of ATCC medium 5 agar).

10.          Incubate the culture at 50-100 µEinsteins/m2/s irradiance at 25°C.  Maintain under a 14/10h light-dark photoperiod.

Name of Depositor RW Krauss
References

Upchurch RG, Elkan GH. Comparison of colony morphology, salt tolerance, and effectiveness in Rhizobium japonicum. Can. J. Microbiol. 23: 1188-1196, 1977. PubMed: 907915

Hwang SW, Hudock GA. Stability of Chlamydomonas reinhardi in liquid nitrogen storage. J. Phycol. 7: 300-303, 1971.

Hwang SW, Horneland W. Survival of algal cultures after freezing by controlled and uncontrolled cooling. Cryobiology 1: 305-311, 1965. PubMed: 5872223

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