久久久久国产成人精品亚洲午夜,成人免费毛片一区二区三区,国产成人久久精品激情91,国产成人无码精品久久久免费,精品欧美国产一区二区三区不卡,国产成人一区二区三区影院,国产精品久久久久久,久久久WWW成人免费精品,欧美91精品久久久久网免费

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.