Paramecium tetraurelia Sonneborn

Handling of test Tube Cultures

This strain is shipped as a growing test tube culture. Upon arrival, remove test tube from sealed plastic envelope, remove plastic seal from cap, loosen the cap one half turn.

1. Add 1.0 ml of ATCC medium 802 bacterized with Enterobacter aerogenes ATCC 13048 twice weekly.  When the tube is filled to within one inch of the top, aspirate from the bottom of the tube and reduce the volume to 5.0 ml.
2. Incubate upright at 25°C with the caps on loosely.

1. Transfer 0.5 ml from a growing culture to 5.0 ml of ATCC medium 802 bacterized with Enterobacter aerogenes (ATCC^®  13048).
2. Add 1.0 ml of ATCC medium 802 bacterized with Enterobacter aerogenes ATCC 13048 twice weekly.  When the tube is filled to within one inch of the top, aspirate from the bottom of the tube and reduce the volume to 5.0 ml.
3. Incubate upright at 25°C with the caps on loosely.

Cryoprotective Solution
DMSO, 1.5 ml
Fresh growth medium w/o bacteria, 7.5 ml
MgCl₂ (0.5 mM), 0.5 ml
CaCl₂ (0.5 mM), 0.5 ml

1. Mix the components in the order listed.  Before adding the MgCl₂ and the CaCl₂ allow the solution to return to room temperature.  When the medium is added to the DMSO the solution will warm up due to chemical heat.
2. Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 200 x g for 1 min.
3. Adjust the concentration of cells to 2 x 10⁵/ml in fresh medium.
4. Mix the cell preparation and the cryoprotective solution in equal portions.
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 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.
7. Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
8. To establish a culture from the frozen state add 1.0 ml ATCC medium 802 to the frozen ampule and place it 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.
9. Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate onto the surface of an ATCC medium 919 (non-nutrient agar) plate containing an overlay of 15.0 ml of bacterized ATCC medium 802.
10. Incubate at 25°C.
11. Once the culture is established, transfer 0.5 ml to 5.0 ml of bacterized ATCC medium 802.
12. Follow the protocol for maintenance of culture.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

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