25%(+/-0% mRNA remaining)(HeLa cell line)
The above cell line was plated in at least triplicate the day before the transfection procedure was started. siRNA was transfected at a final concentration of 100 nM. RNA was extracted from the cells 48 hours after transfection. mRNA levels were measured by real-time RT-PCR using a random primer mix for the RT priming, followed by real time PCR. Knockdown is expressed as the relative amount of target mRNA remaining compared to that of cells transfected with a "nonsense" or scrambled siRNA negative control. Validation results are shown for multiple independent experiments.