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Lipofectamine Plasmid Transfection

594 bytes added, 17:57, 12 August 2009
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[[Category: Transfection]]
==Materials==
*Cells in a 6 well dish, plated and at >90% confluence
*Lipofectamine 2000 (Invitrogen cat# 11668-019 for 1.5mL)
==Protocol (6 well dish)==
#Plate cells the day before so that they are at 90-95% confluence in 2 mL media without antibiotics, or alternatively change media before transfection to OptiMEM
#For each well prepare:
##250uL OptiMEM plus 4 ug of DNA.
##250uL OptiMEM plus 10 uL of Lipofectamine.
##Incubate ~5 minutes at room temperature.
##Combine the two volumes of OptiMEM/DNA and OptiMEM/Lipofectamine.
##Incubate ~20 min at room temperature.
#Add DNA/Lipofectamine complexes (~500 uL) to the 2 mL of media on the cells
#Gently rock plate to mix
#After ~4h re-feed cells with normal media.
Protocol adapted from [http://tools.invitrogen.com/content/sfs/manuals/lipofectamine2000_man.pdf Product Manual]