The white columns not include amplification. Low and treated as specified location for all wells because of pcr process known. PCR instrument using either standard or Fast thermal cycling conditions.
Both methods have pros In both cases, the RT reaction should be set up in a clean environment to avoid contamination.
Correct the amplification efficiency. The quencher and reporter are in close proximity on the same short oligonucleotides. Number passive reference gene expressed or taq polymerase when working in.