Assessment of the assay specificities of the miRCURY and TaqMan platforms. Utilizing the two synthetic samples #1 and #2 with known miRNA contents enabled us to investigate the false positive rate of the individual platforms at given Ct detection thresholds. The rates for the individual platforms are provided in (A) miRCURY and (B) TaqMan. Clearly, the number of false positives increased with increasing Ct detection threshold indicating that carefully chosen Ct detection thresholds could potentially reduce the number of false positives without affecting the true positives. In order to determine the thresholds for the two platforms the raw Ct's of the synthetic samples were compared to no-RT controls. As illustrated by plotting the raw Ct's for the duplicate measurements of the synthetic sample #1 and the no-RT-control for miRCURY (C) and TaqMan (D) thresholds set at Ct = 38 and Ct = 30, respectively would dramatically reduce the number of false positives. #FP, number of false positives.
Jensen et al. BMC Genomics 2011 12:435 doi:10.1186/1471-2164-12-435