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微信小章| 产品名称： | Acanthamoeba griffini Sawyer |
|---|---|
| 商品货号： | TS182671 |
| Strain Designations： | S-7 |
| Application： | Molecular characterization of corneal pathogen |
| 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： | shallow-water, beach-bottom sample, New London, CT, 1962 |
| Product Format： | frozen |
| Type Strain： | no |
| Comments： | species description Group I introns Subgenus systematics based upon SSU rDNA sequence data Molecular characterization of corneal pathogen phylogeny |
| Medium： | ATCC® Medium 997: Fresh water ameba medium |
| Growth Conditions： | Temperature: 25.0°C Duration: grown with bacteria Protocol: ATCCNO: 30011 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in the center of an agar plate of ATCC medium 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days. |
| Subcultivation： | Protocol: ATCCNO: 30011 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in the center of an agar plate of ATCC medium 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days. |
| Cryopreservation： | Cryoprotective Solution DMSO xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 1.5 ml Dryls solution (or similar)xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 8.5 ml 1.xa0xa0xa0xa0 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. xa0xa0 Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryls solution) and washing cells into suspension.xa0 Rub the surface of the plate with a spread bar to detach adhering trophozoites. 3. xa0xa0xa0 Adjust the concentration of cells to at least 2 x 106/ml in fresh medium. 4.xa0 xa0xa0 Mix the cell preparation and the cryoprotective solution in equal portions. 5.xa0 xa0xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6. xa0xa0xa0 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.xa0 Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately -1°C/min.) xa0 7.xa0 xa0xa0 Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator. 8.xa0 xa0xa0 To establish a culture from the frozen state place the vial in a 35°C water bath.xa0 Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0xa0 Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.xa0 Distribute the material evenly over the plate using a spread bar. 9.xa0xa0xa0xa0 Wrap the entire edge of the plate with parafilm and incubate upright at 25°C. Follow the protocol for maintenance of culture. |
| Name of Depositor： | TK Sawyer |
| Chain of Custody： | ATCC < |
| Year of Origin： | 1962 |
| References： | Gast RJ, et al. Discovery of group I introns in the nuclear small subunit ribosomal RNA genes of Acanthamoeba. Nucleic Acids Res. 22: 592-596, 1994. PubMed: 8127708 Sawyer TK. Acanthamoeba griffini, a new species of marine amoeba. J. Protozool. 18: 650-654, 1971. Nerad TA, et al. Acanthamoeba pearcei n. sp. (Protozoa: Amoebida) from sewage contaminated sediments. J. Eukaryot. Microbiol. 42: 702-705, 1995. PubMed: 8520585 Gast RJ, et al. Subgenus systematics of Acanthamoeba: Four nuclear 18S rDNA sequence types. J. Eukaryot. Microbiol. 43: 498-504, 1996. PubMed: 8976608 Ledee DR, et al. Acanthamoeba griffini, molecular characterization of a new corneal pathogen. Invest. Ophthalmol. Vis. Sci. 37: 544-550, 1996. PubMed: 8595954 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 Stothard DR, et al. Fluorescent oligonucleotide probes for clinical and environmental detection of Acanthamoeba and the T4 18S rRNA gene sequence type. J. Clin. Microbiol. 37: 2687-2693, 1999. PubMed: 10405422 Khan NA, et al. Proteases as markers for differentiation of pathogenic and nonpathogenic species of Acanthamoeba. J. Clin. Microbiol. 38: 2858-2861, 2000. PubMed: 10921939 Schroeder JM, et al. Use of subgenic 18s ribosomal dna pcr and sequencing for genus and genotype identification of acanthamoebae from humans with keratitis and from sewage sludge. J. Clin. Microbiol. 39: 1903-1911, 2001. PubMed: 11326011 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 Ledee DR, et al. Advantages of using mitochondrial 16S rDNA sequences to classify clinical isolates of Acanthamoeba. Invest. Ophthalmol. Vis. Sci. 44: 1142-1149, 2003. PubMed: 12601042 Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease in humans. Clin. Microbiol. Rev. 16: 273-307, 2003. PubMed: 12692099 |
| Cross References： | Nucleotide (GenBank) : U07412 nuclear SSU rRNA gene Nucleotide (GenBank) : AF019060 Acanthamoeba hatchetti 2AX1 18S ribosomal RNA gene, partial sequence. Nucleotide (GenBank) : U02540 Acanthamoeba griffini S-7 TS182671 nuclear small subunit ribosomal gene, group I intron. |