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

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      Next entry

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

Methods:

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 https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html

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

Methods:



Personal tools