天天操夜夜拍丨国产精品超清白人精品av丨天天爽天天噜在线播放丨国产精品久久久久久久福利竹菊丨色综合天天天天做夜夜夜夜做丨国内自拍xxxx18丨白人と日本人の交わりビデオ丨午夜激情在线观看丨成人18视频丨久久66热人妻偷产精品丨视频一区二区三区在线观看丨操操操插插插丨久久的久久爽亚洲精品aⅴ丨91精品一区二区三区在线观看丨aaa女人18毛片水真多丨99re在线视频精品丨超碰免费在丨中文字幕人妻无码专区app丨午夜香蕉视频丨又大又粗欧美黑人aaaaa片丨久久夫妻视频丨乌克兰性欧美精品高清丨亚洲欧美福利视频丨少妇高潮久久久久久一代女皇丨97国产高清

eastern oyster, Crassostrea virginica, Cape shore, New Jersey

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.?????????