Colpoda cucullus (Muller) Gmelin (ATCC® 30916)

Organism: Colpoda cucullus (Muller) Gmelin  /  Depositor: DH Lynn

Permits and Restrictions

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Application
testing
Biosafety Level 1
Isolation
dried hay, Toronto, Ontario, Canada, 1970
Product Format dried
Type Strain no
Medium ATCC® Medium 802: Sonneborn's Paramecium medium
Growth Conditions
Temperature: 25.0°C
Protocol: ATCCNO: 30916 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in a T-25 tissue culture flask containing 10 ml ATCC medium 802 inoculated 24 hours previously with Enterobacter aerogenes ATCC 13048. Transfer any residual liquid from the vial to the flask. Place at 25C. Excystment should occur within a few days.
Subcultivation
Protocol: ATCCNO: 30916 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in a T-25 tissue culture flask containing 10 ml ATCC medium 802 inoculated 24 hours previously with Enterobacter aerogenes ATCC 13048. Transfer any residual liquid from the vial to the flask. Place at 25C. Excystment should occur within a few days.
Cryopreservation
1.   Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.

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

3.  While cells are centrifuging prepare a 20% (v/v) solution of sterile DMSO in fresh medium.

a) Add 2.0 ml of DMSO to an ice cold 20 x 150 mm screw-capped test tube;

b) Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 8.0 ml of ice cold medium;

c) Invert several times to dissolve the DMSO;

d) Allow to warm to room temperature.

4.  Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 106 - 107 and 10% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 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. If frozen material is to be stored at temperatures between -130°C and -70°C the shelf life should be empirically tested, i.e., remove stored material at intervals to determine die-off rate.

8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the ampule to a level just above the surface of the frozen material. Do not agitate the ampule.

9. Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate a T-25 tissue culture flask containing 10 ml of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831).

10.          Incubate horizontally with the cap screwed on tightly at 25°C.

Name of Depositor DH Lynn
Year of Origin 1970
References

Wickham SA, Lynn DH. Relations between growth rate, cell size, and DNA content in Colpodean Ciliates (Ciliophora: Colpodea). Eur. J. Protistol. 25: 345-352, 1990.

Nanney DL, et al. Comparison of sequence differences in a variable 23S rRNA domain among sets of cryptic species of ciliated protozoa. J. Eukaryot. Microbiol. 45: 91-100, 1998. PubMed: 9495037

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