国产美女高潮无套久久,无码毛片视频一区二区本码,芒果视频在线观看,亚洲无码午夜免费精品,我和我的老熟女视频,亚洲免费美女动漫精品二区,爆乳放荡的女教师在线观看,国产真人一级a爱做片无码,久久9991电影黄片,日本欧美色三级18禁黄网站,亚洲欧美婬色多多,加勒比熟妇中文高清

eastern oyster, Crassostrea virginica, Cedar Keys, FL, 1998

Temperature: 25.0°C

Duration: axenic

Protocol: ATCCNO: 50439 SPEC: When the frozen ampule arrives, place it directly into a 35C-water bath and transfer its thawed contents to 14 ml of fresh medium in a T-75 tissue culture flask. Maintain by removing 13 ml of cell suspension weekly and replacing with an equal volume of fresh medium. Alternately, aseptically transfer 1.0 ml of a growing culture to 13 ml of fresh medium in a T-75 tissue culture flask. Please note: This strain has wide tolerances to most environmental variables, i.e., temperature range: 15-35C, salinity range: 10-60 parts per thousand; pH range: 6.0-8.5. The distribution of the species worldwide is not known. In order to avert the introduction of this pathogen into non-endemic areas, all culture wastes must be treated as biohazardous and autoclaved prior to disposal. There are no known mechanisms for eradication of this pathogen from the environment.

Protocol: ATCCNO: 50439 SPEC: When the frozen ampule arrives, place it directly into a 35C-water bath and transfer its thawed contents to 14 ml of fresh medium in a T-75 tissue culture flask. Maintain by removing 13 ml of cell suspension weekly and replacing with an equal volume of fresh medium. Alternately, aseptically transfer 1.0 ml of a growing culture to 13 ml of fresh medium in a T-75 tissue culture flask. Please note: This strain has wide tolerances to most environmental variables, i.e., temperature range: 15-35C, salinity range: 10-60 parts per thousand; pH range: 6.0-8.5. The distribution of the species worldwide is not known. In order to avert the introduction of this pathogen into non-endemic areas, all culture wastes must be treated as biohazardous and autoclaved prior to disposal. There are no known mechanisms for eradication of this pathogen from the environment.

1.?? Harvest cells from several cultures which are in logarithmic to late 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 200 x g for 5 min.

4.?? While cells are centrifuging, prepare a 20% solution of DMSO in ATCC Medium 1886.

5.?? Remove the supernatant and pool the cell pellets into a final volume of 4.5 ml.

6.?? Combine the cell suspension with an equal volume of 20% DMSO cryoprotectant solution (prepared in step 4) to yield a final concentration of 10% 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.

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 ATCC medium 1886.

12.Screw the cap on tightly and incubate at 25°C.? Observe the culture daily.? Transfer the culture when many trophozoites are observed.?????????