Transient expression vector - (Aug/03/2009 )
genehunter on Aug 13 2009, 01:38 AM said:
You can select/verify target gene positive clonies from the stable clonies.
Usually i use 30ug of plasmid DNA for transfection but in a 1:1 ratio (15ug from each pBabe and pCMV).
Is this the problem why i'm not getting cells overexpressing the target protein?
Usually you will have different colonies that express at different levels. This is because they vary dramatically in chromosomal integration site and the copy number integrated. You need to do a limited dilution, get the colonies, and screen for individual colonie for protein expression levels.