Real-time PCR was done using 30 ng of human DNA in each of the 96 block positions. (A) A 110 bp amplicon of the beta globin gene was amplified using SYBR Green I detection. (B) The reaction product was also subjected to melting curve analysis. In summary, low variation Cq values (Cq range = 0.16 and SD = 0.033) and overlapping melt curves of the amplicon (TM range of 0.28°C and SD = 0.063) in each of the 96 positions demonstrated temperature homogeneity and equal treatment of all samples - independent of block position.