The CP or cycle threshold was calculated from the amplification plot of the individual reaction graph post the application of 0.07 thresholds. The geometric mean for GAPDH, RPNI, PUM1, B2M, and PMM1 were reported as 15.55, 19.73, 12.22, 18.44, and 21.87 in rectal tumor samples, respectively. The gene discussed is PUM1; the disease is rectal neoplasm.