transfcetion - 6 well plate (Mar/23/2009 )

Hi,

I will start transfection soon to establish stable clones and would like some general guideleines to avoid any confusion. I did this 2 years ago and need to brush up my skills.

I have to transfect 2 different cell lines with a gene construct, a vector control and a mock control. I i use a 6 well plate, do I do it in tripliates, ad that would require 3 plates.

Also, before I start the transfection, is it bests to check the transection efficiency of my 2 different cell lines (both cell lines are normal transformd hepatocytes). I have been told to use a GFP plasmid to check for transfection effiieny? How will I measure the %of efficieny after trasnfectin the cell lines with GFP plasmid?

If your purpose is to establish stable lines, you don't need to do it in triplicates.

Regarding GFP control plasmid, the percent of cells that show GFP under fluorescent microscope is the transfection efficiency.