Difference between revisions of "User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/08/04"

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'''Expand the Thawed [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/08/02 cells]'''
 
'''Expand the Thawed [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/08/02 cells]'''
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Cells became 100% confluent in T-25 flask.  
 
Cells became 100% confluent in T-25 flask.  
 
wash the cells with 2 mL 1xPBS, trypsinize them with 0.5 mL of Trypsin buffer and transfer the entire culture into a T-75 flask with 10 mL total growth medium (10% FBS, 1% pen-strep). After the t-75 is 100% confluent, you can make new frozen stocks or passage the cells at 1:10 (or 1:5 if they are slow-growing).
 
wash the cells with 2 mL 1xPBS, trypsinize them with 0.5 mL of Trypsin buffer and transfer the entire culture into a T-75 flask with 10 mL total growth medium (10% FBS, 1% pen-strep). After the t-75 is 100% confluent, you can make new frozen stocks or passage the cells at 1:10 (or 1:5 if they are slow-growing).

Revision as of 09:37, 5 August 2013

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08/04/2013

Expand the Thawed cells

Cells became 100% confluent in T-25 flask. wash the cells with 2 mL 1xPBS, trypsinize them with 0.5 mL of Trypsin buffer and transfer the entire culture into a T-75 flask with 10 mL total growth medium (10% FBS, 1% pen-strep). After the t-75 is 100% confluent, you can make new frozen stocks or passage the cells at 1:10 (or 1:5 if they are slow-growing).