I am trying to quantify archaea by q-pcr. First I wanted to see how my standarts work . I used 6 standarts I obtained a standart curve at the end of the reaction. I wanted to decrease the error by discarding some of the standarts from calculation. I eliminated two of the standarts. However after I discarded some of the standarts the crossing points of the included ones changed. How could it be possible? I did the same thing before in another q-pcr machine, CP values didn't change. By the way I started to use Light Cycler 480.