原核和真核细胞表达系统课件.ppt

Procedure of monolayer sub culture Monolayer should reach to confluency in 2-4 days. Serum supplemented cultures do not adhere to surface tightly where as serum free attach very tightly to substrates Aspirate medium floating cells from a confluent monolayer discard them. Add 4ml of RT complete growth medium to each 25cm2 flask(12 ml to a 75 cm2 flask) Resuspend cells by pipetting the medium across the monolayer with a Pasteur pipette. (Enzymatic dissociation is not recommended) Observe cell monolayer using an inverted microscope to ensure adequate cell detachment Perform viable cells count o