1 reply to this topic

[Mariavi]

Hello,

I have recently used Sigma esiRNA against SAT1 gene, and I was very surprissed when I checked the transfections using Real Time PCR and discovered that, in trhee independent experiments, cells treated with siRNA for 24h showed more mRNA levels than non transfected cells; the universal negative control also induced expression although not to the same extent. I insist, this happened in three independent experiments.

Other member of my research group used Sigma esiRNA against another gene and he had a mRNA induction too. He also checked de PCR products in an agarose gel, and the bands showed that transfected cells had more mRNA than controls.  

I wonder if someone has ever had some problems like this, and if there is any explanation and solution.

Thank you in advance for your help.

[pcrman]

This is weired, though I have often heard that people see increase in gene expression while expecting RNAi. Can you confirm the increase in mRNA expression by Western? Why don't you just use a single siRNA for your target gene.