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Measure transfection effiicency - referee's question - (Dec/06/2011 )

I am trying to get my paper accepted but the referee has asked me to verfify that my construct does not have some "special property" that enables its more rapid uptake into my cells that the Renilla control. He suggested qRT-PCR for the vector backbone.

I have designed primers for the ampicillin resistance gene but how to I normalise for differences in cell numbers, as you dont have an endogenous reference like with cDNA expression.

Any thoughts would be great.


why not lift and count the cells, then lyse in your RNA extraction buffer at a defined number per ml?