I performed a qPCR experiment using QIAGEN's miRCURY LNA RT kit and miRCURY LNA SYBR Green Kits to investigate the miRNA expression levels in some genes, but I could not obtain the desired result in the melting curve analysis. I conducted the experiment on a Roche LightCycler 480-II, and I used U6 as a reference gene. Actually, the Ct values ​​and melting curve I obtained for U6 are not so bad, but unfortunately I am facing this problem in some of the miRNAs. The RNA source I used is of FFPE origin and I assume that the quality of some RNA samples is relatively not very high. Since I consider this situation, I diluted the cDNAs as 1:7 after the reverse transcription step. However, before using this kit I had efficient results by using TAKARA and miScript kits and properly optimizing them. I should also point out that although I have also ordered QIAGEN's own designed primer assays and carefully followed the steps in the protocol, why am I not getting an effective result from the qPCR experiment? What would you suggest in order to get rid of non-specific PCR products? I would be very grateful if there are some researchers who could share their opinions and supports on performing their experiment by using the miRCURY LNA SYBR Green kit.