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siRNA - augmentation instead of knockdown - siRNA - augmentation instead of knockdown (Apr/02/2005 )

I have tried to do knockdown to Luciferase gene using luciferase reporter plasmid in HEK293 cells and OligonucleotideLuciferase provide to me in the kit "pFIV-H1-copGFPT siRNA Cloning and Expression Vector".
I used 3ug of luciferase reporter plasmid and 10 ug of the siRNA using CaCl2 transfection.
After 48 hours the cells were observed under the fluorescent microscope (transfection effeciency was >80%), but when i monitored luciferase activity the measurements without siRNA were - 6000 (arbitrary units), with non related siRNA in the FIV vector -8000 and with siRNA OligonucleotideLuciferase -12,000
The results repeated twice.

I would like to understand why did i get augmentation of the signal instead of knockdown of the gene.
What was wrong?

Thanks

-luc-

Where does your siRNA target to? Have you checked there is any homology, even partial, to other genes and non-coding regions?

-pcrman-

Molar ratio of Luc reporter to siRNA vector is a key. Make sure each cell getting Luc also getting sufficient siRNA......

-postdoc2130-