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微信小章| 产品名称： | Cercomonas vibrans Sandon |
|---|---|
| 商品货号： | TS145317 |
| Deposited As： | Cercobodo sp. |
| Biosafety Level： | 1
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： | dry soil, Mexico City, Mexico |
| Product Format： | test tube |
| Type Strain： | no |
| Comments： | This is the only species of Cercomonas known to feed on other protists. |
| Medium： | ATCC® Medium 1967: Soil freshwater cereal grass medium |
| Growth Conditions： | Temperature: 25.0°C Protocol: This strain is shipped as a growing culture and includes Adriamonas sp. (food source for Cercomonas vibrans) and Enterobacter aerogenes ATCC 13048 (food source for Adriamonas sp.). The food will not be completely eliminated by the predator, C. vibrans. The Adriamonas sp. will eventually encyst. C. vibrans also produces cysts. Because those cysts are not very stable, the culture must be passaged every 7-10 days to ensure the cultures continued viability. Upon arrival of the shipment, agitate the culture, aseptically transfer the entire contents to a T-25 tissue culture flask, and incubate it at 25C. The bacterial flora present in the shipped culture will support growth of the Adriamonas sp., but growth can be improved by using medium that has been bacterized with E. aerogenes ATCC 13048 approximately 24 hours before inoculation with the Cercomonas/Adriamonas culture. (Other species of bacteria may work equally well, but this one has been empirically assessed.) After bacterization of the medium, agitate the culture of TS145317 and aseptically transfer 0.25 ml to the new medium. Screw the cap on tightly and incubate at 25C. |
| Subcultivation： | Protocol: This strain is shipped as a growing culture and includes Adriamonas sp. (food source for Cercomonas vibrans) and Enterobacter aerogenes ATCC 13048 (food source for Adriamonas sp.). The food will not be completely eliminated by the predator, C. vibrans. The Adriamonas sp. will eventually encyst. C. vibrans also produces cysts. Because those cysts are not very stable, the culture must be passaged every 7-10 days to ensure the cultures continued viability. Upon arrival of the shipment, agitate the culture, aseptically transfer the entire contents to a T-25 tissue culture flask, and incubate it at 25C. The bacterial flora present in the shipped culture will support growth of the Adriamonas sp., but growth can be improved by using medium that has been bacterized with E. aerogenes ATCC 13048 approximately 24 hours before inoculation with the Cercomonas/Adriamonas culture. (Other species of bacteria may work equally well, but this one has been empirically assessed.) After bacterization of the medium, agitate the culture of TS145317 and aseptically transfer 0.25 ml to the new medium. Screw the cap on tightly and incubate at 25C. |
| Cryopreservation： | Cryoprotective Solution DMSO xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 2.0 ml Fresh growth medium w/o bacteriaxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 8.0 ml 1.xa0xa0xa0xa0 Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat. 2. xa0xa0 Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min. 3.xa0 Adjust the concentration of cells at least 2 x 106/ml in fresh medium. 4.xa0 xa0xa0 Mix the cell preparation and the cryoprotective solution in equal portions. 5.xa0 xa0xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6. xa0xa0xa0 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.xa0 Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately -1°C/min.) xa0 7.xa0 xa0xa0 Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator. 8.xa0 xa0xa0 To establish a culture from the frozen state place the vial in a 35°C water bath.xa0 Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0xa0 Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 ml ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048). 9.xa0xa0xa0xa0 Incubate at 25°C with the cap screwed on tightly. 10.xa0xa0 Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 ml to 10.0 ml of bacterized ATCC medium 802. 11.xa0xa0 Follow the protocol for maintenance of culture. |
| Name of Depositor： | J Martinez-Cruz |
| Chain of Custody： | ATCC < |
| References： | Jovita Martinez-Cruz, personal communication |