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Naegleria gruberi Schardinger
Naegleria gruberi Schardinger
規(guī)格:
貨期:
編號(hào):B210004
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(chēng) Naegleria gruberi Schardinger
商品貨號(hào) B210004
Strain Designations CCAP 1518/1g
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 Freshwater, Tuskegee, AL, 1965
Product Format dried
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
Comments
Zymodeme Gr-10
Maximum temperature tolerance 37°C
Equivalent to CCAP 1518/1g, maximum temperature tolerance 36°C
Biochemical identification
Genetic cluster E [Int. J. Parasitol. 19: 823-834, 1989] = Naegleria gruberi unknown cluster [J. Eukaryot. Microbiol. 40: 179-187, 1993].
Equivalent to CCAP 1518/1g, Group I intron in SSUrRNA gene
Medium ATCC® Medium 997: Fresh water ameba medium
ATCC® Medium 711: PYB
Growth Conditions
Temperature: 20°C to 25°C
Cryopreservation Reagents
Cryoprotective Solution
DMSO, 1.5 mL
Page's Balanced Salt Solution (or similar), 8.5 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 which is at or near peak density by adding 5 mL ATCC medium 1323 (Page's Balanced Salt Solution) and washing cells into suspension.  Rub the surface of the plate with a spread bar to detach adhering trophozoites.
  3. Adjust the concentration of cells to at least 2 x 106/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. 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.)  
  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°C water bath.  Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.   Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.  Distribute the material evenly over the plate using a spread bar.
  9. Wrap the entire edge of the plate with parafilm and incubate upright at 20-25°C.
  10. Follow the protocol for maintenance of culture.
Name of Depositor CCAP
Chain of Custody
ATCC <-- CCAP <-- F.C. Page 48
Year of Origin 1965
References

Page FC. Taxonomic criteria for limax amoebae, with descriptions of 3 new species of Hartmannella and 3 of Vahlkampfia. J. Protozool. 14: 499-521, 1967. PubMed: 6050658

Daggett PM, Nerad TA. The biochemical identification of vahlkampfiid amoebae. J. Protozool. 30: 126-128, 1983. PubMed: 6864593

Page FC. Morphological variation in the cyst wall of Naegleria gruberi (amoebida, vahlkampfiidae). Protistologica 11: 195-204, 1975.

De Jonckheere JF. A Group I intron in the SSUrDNA of some Naegleria spp. demonstrated by polymerase chain reaction amplification. J. Eukaryot. Microbiol. 40: 179-187, 1993. PubMed: 8461891

Adams M, et al. A genetic approach to species criteria in the amoeba genus Naegleria using allozyme electrophoresis. Int. J. Parasitol. 19: 823-834, 1989. PubMed: 2635158

Robinson BS, et al. Discontinuous genetic variation among mesophilic Naegleria isolates: Further evidence that N. gruberi is not a single species. J. Protozool. 39: 702-712, 1992. PubMed: 1453360

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