| 产品名称： |
Sapocribrum chincoteaguense |
| 商品货号： |
TS165704 |
| Deposited As： |
Sexangularia sp. |
| Strain Designations： |
Chinc5 |
| 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： |
Moist soil from mud flat approximately 800 yards from the ocean, Chincoteague, VA, 2002 |
| Product Format： |
frozen |
| Storage Conditions： |
Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures:
2-8°C
Live Cultures:
See Protocols section for handling information |
| Type Strain： |
no |
| Medium： |
ATCC® Medium 1525: Seawater 802 medium
|
| Growth Conditions： |
Temperature: 25°C
Culture System:xa0Grown with Enterobacter aerogenesxa0(ATCC® 13048™) and mixed bacteria |
| Cryopreservation： |
Reagents
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium w/o bacteria, 8.0 mL
Harvest and Preservation
- Mix the components in the order listed. When the medium is added to the DMSO, the solution will warm up due to chemical heat.
- Harvest cells from a culture that is at or near peak density by centrifugation atxa0800 x g for 5 min.
- Adjust the concentration of cells to at least 2 x 107/mL in fresh medium.
- Mix the cell preparation and 20% (v/v) DMSO in equal portions. The final concentration will be
1xa0x 107 cells/mL and 10% DMSO. The time from the mixing of the cell preparation andxa0cryoprotective solution to the start of the freezing process should be no less than 15 min. andxa0no more than 60 min.
- Dispense in 0.5 mL aliquots into 1.0 - 2.0xa0mL sterile plastic screw-cappedxa0cryules (specialxa0plastic vials for cryopreservation).
- 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,xa0place 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 thisxa0apparatus is approximately -1°C/min.)
- Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
- To establish a culture from the frozen state, place the vial in a 35°C water bath. Immersexa0the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0Immediately after thawing, do not leave in water bath; aseptically remove the contents of thexa0ampule and inoculate into a T-25xa0tissue culture flask containing 10 mL ATCC medium 1525xa0bacterized with Enterobacter
aerogenes (ATCC® 13048™).
- Incubate at 25°C with the cap screwed on tightly.
- Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 mLxa0to 10.0 mL of bacterized ATCC medium 1525.
- Follow the protocol for maintenance of culture.
|
| Name of Depositor： |
TA Nerad |
| References： |
Thomas A Nerad, personal communication
Lahr DJ, et al. Sapocribrum chincoteaguense n. gen. n. sp.: A Small, Scale-bearing Amoebozoan with Flabellinid Affinities. J Eukaryot Microbiol doi: 10.1111/jeu.12199 Epub ahead of print, 2014. PubMed: 25515047
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