Figure 1

Calibration with the exogenous control cel-miR-39-3p corrects for a majority of technical assay variance. (A) Within each study participant (n = 22) (i) the median raw Cq values of the 36 always-observed miRNAs (solid grey line), (ii) mean raw Cq values for cel-miR-39-3p control (solid black line), and (iii) the average of the median Cq values of miRNAs always observed and cel-miR-39-3p (dashed grey line) are shown for each visit number. Spearman rank correlation was used to relate the median Cq values of the 36 miRNAs always observed to mean Cq values for cel-miR-39-3p (r = 0.89). (B) Within each study participant (i) the mean raw Cq values of miR-16-5p, the primary endogenous miRNA used for normalization (solid grey line) and (ii) mean raw Cq values for cel-miR-39-3p (solid black line) are shown for each visit number. Spearman rank correlation was used to relate the mean Cq values of miR-16-5p to those of cel-miR-39-3p (r = 0.92). (C) Within each study participant the raw Cq values were calibrated to cel-miR-39-3p values. The calibrated mean Cq values for miR-16-5p (solid grey line) and cel-miR-39-3p (solid black line) for each visit number are shown. The mean residual Spearman rank correlation (r = 0.02) is the correlation that remains after calibration is applied. In panels (B) and (C) the qPCR assay technical replicates for miR-16-5p and cel-miR-39-3p are depicted by the circles.