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Acanthamoeba pearcei Nerad et al. 拉丁名

(ATCC® 50435™) 統(tǒng)一編號

Strain Designations 菌種別名 205-1

Biosafety Level 生物安全等級 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 分離基物 seabottom sample 50 meters deep (38 degrees 19.6'N, 74 degrees 18.0'W) at a site used for disposal of barge-delivered wastes from 1973-1980, Atlantic Ocean approximately 65 km seaward from the coast of Delaware and Maryland, 1978

Product Format 提供形式 dried

Type Strain 模式菌株 yes

Medium 培養(yǎng)基 ATCC® Medium 997: Fresh water ameba medium

Growth Conditions 生長條件

Temperature 培養(yǎng)溫度: 25.0℃

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 1. Allow the cells to encyst. To detach cysts from the plate flush the surface with 5 ml fresh ATCC medium 1323 (Page's Balanced Salt Solution). Rub the surface of the plate with a spread bar to detach adhering cysts.

2. Transfer the liquid medium to a sterile centrifuge tube.

3. If the cyst concentration does not exceed 2 x 106 cysts/ml adjust the suspension to that concentration. To adjust the concentration, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield 2 x 106.

4. 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.

5. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be at least 106 cysts/ml and 7.5% (v/v) DMSO. The equilibration time (the time between addition of DMSO and the start of the cooling cycle) should be no less than 15 min and no longer than 30 min.

6. Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

7. Place the vials in a controlled rate freezing unit. From room temperature cool at -1℃/min to -40℃. If the freezing unit can compensate for the heat of fusion, maintain rate at -1℃/min through the heat of fusion. At -40℃ plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1℃ freezing apparatus. 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.)

8. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer. 9. To establish a culture from the frozen state place an ampule in a water bath set at 35℃ (2-3 min). Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.

10. Immediately after thawing, aseptically remove the contents of the ampule and distribute to the center of a fresh plate of ATCC medium 7

11. Distribute the material evenly over the plate using a spread bar. Incubate at 25℃.

Name of Depositor 寄存人 TK Sawyer

Year of Origin 收藏年份 1973

References 參考文獻 Nerad TA, et al. Acanthamoeba pearcei n. sp. (Protozoa: Amoebida) from sewage contaminated sediments. J. Eukaryot. Microbiol. 42: 702-705, 1995. PubMed: 8520585

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

type strain

Gast RJ. Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. J. Eukaryot. Microbiol. 48: 609-615, 2001. PubMed: 11831768

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) : AF019053 Acanthamoeba pearcei 18S ribosomal RNA gene, partial sequence.