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Acanthamoeba polyphaga (Puschkarew) Page
Acanthamoeba polyphaga (Puschkarew) Page
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貨期:
編號:B190381
品牌:Mingzhoubio

標準菌株
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DNA
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產品名稱 Acanthamoeba polyphaga (Puschkarew) Page
商品貨號 B190381
Strain Designations CCAP 1501/3a
Application
characterization of Acanthamoeba polyphaga
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, Drummond, WI, 1964
Product Format frozen
Type Strain no
Comments
Re-definition of the genus
mitochondrial DNA fingerprinting
Restriction fragment length polymorphisms of DNA
phylogeny
characterization of Acanthamoeba polyphaga
Medium ATCC® Medium 712: PYG w/ Additives
Growth Conditions
Temperature: 25.0°C
Growth condition: Axenic
Protocol: This strain is distributed as a frozen preparation. See the general procedures for thawing a frozen vial. Once vial is thawed, aseptically transfer the entire contents to a 16 X 125 mm screw-capped test tube or a T-25 tissue culture flask containing 5.0 ml of ATCC medium 712. Incubate horizontally at 25C. Trophozoites should be evident in 1-5 days.
Subcultivation
Protocol: This strain is distributed as a frozen preparation. See the general procedures for thawing a frozen vial. Once vial is thawed, aseptically transfer the entire contents to a 16 X 125 mm screw-capped test tube or a T-25 tissue culture flask containing 5.0 ml of ATCC medium 712. Incubate horizontally at 25C. Trophozoites should be evident in 1-5 days.
Cryopreservation

1.?? To achieve the best results set up cultures with several different inocula (e.g. 0.25 ml, 0.5 ml, 1.0 ml).? Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density.

2.? If the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107cysts/ml with fresh medium.? If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.

3.? While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows:? Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.? Allow the DMSO to solidify.? Add the required volume of refrigerated medium.? Dissolve the DMSO by inverting the tube several times.?

????? *NOTE: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.

4.? Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/ml and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 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 the vials in a controlled rate freezing unit.? From room temperature cool at -1°C/min to -40°C.? If the freezing unit can compensate for the heat of fusion, maintain rate at??????? -1°C/min through the heat of fusion.? At -40°C plunge 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. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.

8.?? To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.

9.?? Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 ml of fresh ATCC medium 712 in a T-25 tissue culture flask or plastic 16 x 125 mm screw-capped test tube.? Incubate at 25°C.

Name of Depositor CCAP
Chain of Custody
ATCC <<--CCAP<<--F.C. Page 23
Year of Origin 1964
References

Page FC. Re-definition of the genus Acanthamoeba with descriptions of three species. J. Protozool. 14: 709-724, 1967. PubMed: 5604481

McLaughlin GL, et al. Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae. J. Clin. Microbiol. 26: 1655-1658, 1988. PubMed: 2903176

Gautom RK, et al. Mitochondrial DNA fingerprinting of Acanthamoeba spp. isolated from clinical and environmental sources. J. Clin. Microbiol. 32: 1070-1073, 1994. PubMed: 7913095

Daggett PM, et al. Distribution and possible interrelationships of pathogenic and nonpathogenic Acanthamoeba from aquatic environments. Microb. Ecol. 8: 371-386, 1982.

Daggett PM, et al. A molecular approach to the phylogeny of Acanthamoeba. Biosystems 18: 399-405, 1985. PubMed: 4084681

Fritsche TR, et al. Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J. Clin. Microbiol. 31: 1122-1126, 1993. PubMed: 8501212

Stothard DR, et al. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J. Eukaryot. Microbiol. 45: 45-54, 1998. PubMed: 9495032

Kong HH, et al. Interstrain polymorphisms of isoenzyme profiles and mitochondrial DNA fingerprints among seven strains assigned to Acanthamoeba polyphaga. Korean J. Parasitol. 33: 331-340, 1995. PubMed: 8591011

Kong HH, et al. Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J. Clin. Microbiol. 40: 1199-1206, 2002. PubMed: 11923331

Alves JMP, et al. Random amplified polymorphic DNA probes as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba. Braz. J. Med. Biol. Res. 33: 19-26, 2000.

Khan NA. Pathogenicity, morphology, and differentiation of Acanthamoeba. Curr. Microbiol. 43: 391-395, 2001. PubMed: 11685503

Khan NA, Paget TA. Molecular tools for speciation and epidemiological studies of Acanthamoeba. Curr. Microbiol. 44: 444-449, 2002. PubMed: 12000996

Khan NA, Tareen NK. Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains. Folia Parasitol. 50: 97-104, 2003. PubMed: 14560969

Cross References

Nucleotide (GenBank) : AF019061 Acanthamoeba polyphaga Page-23 18S ribosomal RNA gene, partial sequence.

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李經理 13626845108 972239479
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