I did a cDNA synthesis using 500ng of RNA with helps of RevertAid enzyme from ThermoScientific. The result is quite surprising, because from the electrophoresis of PCR product, I concluded that there was a DNA contamination. The primer I used was designed to skip 1 intron, so amplified region with DNA samples would be much longer than the RNA samples.
After I used a different kind of RNA isolation method, which was using TRIZOL, I got less DNA contamination but the band of DNA contamination was still more significant than the cDNA result according to electrophoresis of PCR Product.
I wonder if I have to add more RNA to the mixture. What is your experience? Please kindly share.