User:Cassandra M Barrett/Notebook/Haynes Lab/2015/09/06

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Plating Mammalian Cells

Purpose: to finish TC training with Rene by plating cells

1:10 dilution flask and 6well plate were created from flask with 90% confluence


Calculate how much of 90% confluent culture to add to media so that you will get about 0.3X10^6 cells per well in 4mL of media Use M1V1=M2V2

Check cell density in flask with scope Aspirate out media Wash with 3mL PBS Aspirate PBS Add 1mL trypsin Allow cells to detach and add 9mL media (DMEM+FBS+Antibiotic)

Start new 750 Flask with 9mL of fresh media and 1mL of culture

Aliquot 20.4mL media into a tube and add 3.6mL of culture Aliquot 4mL per well

Incubate in CO2 incubator