Derek Houston Already tried these things. Self-binding is not an issue. No significant secondary structures were found by using online oligoanalyzer tools.
Jetty Ramadevi Yes, I'm on it. Gradient at 59.1 degree Celsius is displaying the best outcome but not for my every sample. I've now tried extracting DNA from a different technique to maintain the concentration of my samples equally. The results are not that consistent. It should have displayed a common allelic band in every sample, post-amplification.