User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/06/17: Difference between revisions
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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 2mL Trypsin to each of the wells 1 and 2 for 4 minutes. | ||
*Add 3.5mL media to neutralize Trypsin. | *Add 3.5mL media to neutralize Trypsin. | ||
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*Take 1mL of cells in well 2, dilute with 9mL of media and transfer to 10cm plate 1x10<suP>4</sup>. | *Take 1mL of cells in well 2, dilute with 9mL of media and transfer to 10cm plate 1x10<suP>4</sup>. | ||
*Take 0.5mL of cells in well 2, dilute in 9.5mL of media and transfer to 10cm plate 1x10<suP>3</sup>. | *Take 0.5mL of cells in well 2, dilute in 9.5mL of media and transfer to 10cm plate 1x10<suP>3</sup>. | ||
** Keep cells in the incubator to reach 90% confluency. | |||
Revision as of 16:04, 23 July 2013
 PcTF Subcloning in E. coli
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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
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