日韩精品人妻一区二区中文,五月婷婷综合在线视频,A久久精品国产精品亚洲,亚州中文精品有码视频在线,精品1区2区3区产品乱码,色哟哟免费观看视频入口,婷婷射精AV这里只有精品,2024日产乱码国产,国产精品三级一区二区,艳妇乳肉豪妇荡乳AV

eastern oyster, Crassostrea virginica, LA, Gulf Coast

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.?????????

ATCC <<--D Bushek<<--J. LaPeyre