波多野结衣毛片-乳色吐息在线观看-国产伦子伦对白视频-性做久久久-狠狠干2019-黄色裸体片-美女无遮挡免费网站-国产91熟女高潮一区二区-懂色av蜜臀av粉嫩av分享-小h片在线观看-台湾佬在线-日韩激情在线播放-欧日韩不卡在线视频-波多野结衣中文字幕一区-天天操夜夜草

Certified Reference Material produced under an ISO 17034 accredited process.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca²⁺/Mg²⁺ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.025% (w/v) Trypsin-EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 1.0 to 2.0 mL of 0.025% Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1.5 X 10⁴ to 3.0 X 10⁴ viable cells/cm^2 is recommended.
7. Incubate cultures at 37°C.