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Trypanosoma cruzi Chagas
Trypanosoma cruzi Chagas
規(guī)格:
貨期:
編號(hào):B243449
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Trypanosoma cruzi Chagas
商品貨號(hào) B243449
Strain Designations CA-I CL72 Lampit Resistant
Application
Vector borne research
Biosafety Level 2

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
Lampit (=Nifurtimox) resistant strain derived from CA-I CL72 (=ATCC 50791)
Product Format frozen
Storage Conditions Frozen: -70°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
Axenic/Xenic Axenic
Type Strain no
Medium ATCC® Medium 1029: LIT medium
Growth Conditions
Temperature: 20-25°C
Cryopreservation
  1. Harvest cells from several cultures in very late logarithmic to early stationary phase of growth.  Vigorously agitate to suspend the cells.
  2. Aseptically transfer the cell suspension to 15 ml plastic centrifuge tubes.
  3. Centrifuge at ~800 x g for 5 min.
  4. While cells are centrifuging, prepare a 10% solution of DMSO in complete ATCC Medium 1029.  Cool on ice.
  5. Remove the supernatant and pool the cell pellets to the final volume desired with fresh growth medium.
  6. Combine the cell suspension with an equal volume of 10% DMSO cryoprotectant solution (prepared in step 4) to yield a final concentration of 5% DMSO.
  7. Dispense in 0.5 ml aliquots to 1.0-2.0 ml Nunc vials (special plastic vials for cryopreservation).
  8. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  At -40°C, plunge ampules 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.).  
  9. Store ampules in a liquid nitrogen refrigerator until needed.
  10. To establish a culture from the frozen state, place a frozen ampule in a 35°C water bath just enough to cover the frozen material.  Allow the ampule to thaw completely (2-3 min).
  11. Immediately after thawing, aseptically remove the contents and transfer to a T-25 tissue culture flask containing 10 ml of fresh complete ATCC medium 1029.
  12. Screw the cap on tightly and incubate at 20-25°C.    Observe the culture daily and transfer when numerous trophozoites are observed.         
Name of Depositor JA Dvorak
Special Collection NCRR Contract
References

Nozaki T, et al. Cellular and molecular biological analyses of nifurtimox resistance in Trypanosoma cruzi. Am. J. Trop. Med. Hyg. 55: 111-117, 1996. PubMed: 8702014

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