Sapocribrum chincoteaguense 菌株拉丁名
(ATCC® 50979™) 統(tǒng)一編號(hào)
Deposited As Sexangularia sp.
Strain Designations 菌株別名 Chinc5
Biosafety Level 生物安全等級(jí) 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 分離源 Moist soil from mud flat approximately 800 yards from the ocean, Chincoteague, VA, 2002
Product Format 提供形式 frozen
Storage Conditions 保藏方法
Frozen Cultures 冷凍培養(yǎng) : -70℃ for 1 week; liquid N2 vapor for long term
storage 保藏方法 Freeze-dried
Cultures: 2-8℃
Live Cultures 活菌 : See Protocols section for handling information
Type Strain 模式菌株 no
Medium 培養(yǎng)基 ATCC® Medium 1525: Seawater 802 medium
Growth Conditions 生長條件
Temperature 培養(yǎng)溫度 : 25℃
Culture System: Grown with Enterobacter aerogenes (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
1.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.Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.
3.Adjust the concentration of cells to at least 2 x 107/mL in fresh medium.
4.Mix the cell preparation and 20% (v/v) DMSO in equal portions. The final concentration will be 1 x 107 cells/mL and 10% DMSO. The time from the mixing of the cell preparation and cryoprotective solution to the start of the freezing process should be no less than 15 min. and no more than 60 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 vials in a controlled rate freezing unit. From room temperature, cool at -1℃/min to -40℃. If freezing unit can compensate for the heat of fusion, maintain rate at -1℃/min through heat of fusion. At -40℃, plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1℃ freezing apparatus.
6.Place the apparatus at -80℃ for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1℃/min.)
7. Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
8.To establish a culture from the frozen state, place the vial in a 35℃ water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. 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 1525 bacterized with Enterobacter aerogenes (ATCC® 13048™).
9.Incubate at 25℃ with the cap screwed on tightly.
10.Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 mL to 10.0 mL of bacterized ATCC medium 1525.
11.Follow the protocol for maintenance of culture.
Name of Depositor 寄存人 TA Nerad
References 參考文獻(xiàn) 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
