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==Protocol==
#Warm OptiMEM, COS-FBS Media and PBS -/-.
#Calculate amount of Fugene needed.
*Per ug of DNA need 3 uL Fugene.
*Per uL of Fugene need 16 uL of OptiMEM.
#Add Fugene to OptiMEM, incubate ~5 min.
#Add required amount of DNA to eppendorf tubes.
#Add 51 uL/ug OptiMEM/Fugene to each DNA tube.
#Incubat 5-20 min.
#Split confluent cells 2-3X into fresh dishes as follows:
#Wash confluent COS cell plate(s) twice with 10 mL D-PBS -/-
#Add 1 mL Trypsin solution (0.05%) and incubate at 37C until cells are detached
#Add 25 mL COS/FBS
#Aliquot cells into wells as required (1 mL per 12well, 2 mL per 6 well)
#Add DNA/Fugene/DMEM to cells
#Leave mixture on Cells for 24-48h to allow protein to accumulate
#Warm OptiMEM, COS-FBS Media and PBS -/-.
#Calculate amount of Fugene needed.
*Per ug of DNA need 3 uL Fugene.
*Per uL of Fugene need 16 uL of OptiMEM.
#Add Fugene to OptiMEM, incubate ~5 min.
#Add required amount of DNA to eppendorf tubes.
#Add 51 uL/ug OptiMEM/Fugene to each DNA tube.
#Incubat 5-20 min.
#Split confluent cells 2-3X into fresh dishes as follows:
#Wash confluent COS cell plate(s) twice with 10 mL D-PBS -/-
#Add 1 mL Trypsin solution (0.05%) and incubate at 37C until cells are detached
#Add 25 mL COS/FBS
#Aliquot cells into wells as required (1 mL per 12well, 2 mL per 6 well)
#Add DNA/Fugene/DMEM to cells
#Leave mixture on Cells for 24-48h to allow protein to accumulate