I know that my target is in my sample as I extracted it from cell culture myself. These plots show some amplification. The first one seems O.K for the first 36 cycles but then collapses at the end. Do you think that suggests that the target is being picked up but it's not quite working as it should. I've being using SYBR with no ROX. I'm guessing that maybe using SYBR with ROX or a passive reference of some sort might be the way to go. Help much appreciated. Thanks.