User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/06/17: Difference between revisions
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= | =06/17/2013= | ||
''' | '''Prepare Transfected Cells for Hygromycine Treatment''' | ||
Transfer [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/06/10 Transfected Cells] from 6 well plate to 10cm cell culture dishes. | |||
Target cell concentrations are: 1x10<suP>6</sup>, 1x10<suP>5</sup>, 1x10<suP>4</sup>, 1x10<suP>3</sup> | |||
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'''1x10<suP>6</sup> Monolayer 100% confluent cells are attached to the bottom of each well ''' | |||
*Add 2mL Trypsin to each of the wells 1 and 2 for 4 minutes. | |||
*Add 3.5mL U2OS media (p/s) to neutralize Trypsin. | |||
*Add 6mL U2OS media (p/s) to the 4ml of well 1. | |||
*Transfer all 10mL of well 1 to 10cm plate to have 1x10<suP>6</sup>. | |||
*Add 16mL U2OS media (p/s) to the 4mL of well 2. | |||
*Take 2mL of cells, dilute with 8mL of U2OS media (p/s) and transfer to 10cm plate 1x10<suP>5</sup>. | |||
*Take 1mL of cells in well 2, dilute with 9mL of U2OS media (p/s) and transfer to 10cm plate 1x10<suP>4</sup>. | |||
*Take 0.5mL of cells in well 2, dilute in 9.5mL of U2OS media (p/s) and transfer to 10cm plate 1x10<suP>3</sup>. | |||
'' Keep cells in the incubator to reach 90% confluency.'' |
Revision as of 16:15, 23 July 2013
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06/17/2013Prepare Transfected Cells for Hygromycine Treatment Transfer Transfected Cells from 6 well plate to 10cm cell culture dishes. Target cell concentrations are: 1x106, 1x105, 1x104, 1x103 1x106 Monolayer 100% confluent cells are attached to the bottom of each well
Keep cells in the incubator to reach 90% confluency. |