I am trying to determine DNA copy number in patient peripheral blood after infusion of transduced cells. I am getting a lot of background in early cycles and in negative samples, I have tried to set the baseline higher and played with the threshold but it doesn't seem to help and I am getting CTs of 2 which is not right!! I am using Taqman, an ABI Step One plus machine and multipex VIC/FAM probes quenched with TAMRA. The samples are lysed in SDS/PK from frozen cells.
qPCR copy number analysis