Predictions of probe behaviors at 48°C hybridization temperature. Set 1 (red lines) and Set 2 (green lines) calibration results as functions of target sample concentration. (a) Calibrated target intensities (solid lines) and their regression equations (dotted lines) at 48°C. The horizontal blue line limits intensity-to-concentration reverse estimates to the linear range. (b) Target-to-nontarget intensity ratios lines. The blue line represents estimated experiment errors attributable to nontarget background differences; note that it is in linear scale.