Assessment of the platforms ability to discriminate miRNAs with close sequence homology. Using the synthetic samples, with known miRNA content, it was investigated how many of the false positives detections (miRNAs not in the sample) at a given Ct detection threshold could be related to sequence homology to a miRNA in the sample. Plotted are the number of false positives detected in the synthetic samples at given Ct detection thresholds and it is indicated how many of these can be related to sequence homology. (A, B) TaqMan analysis of synthetic samples #1 and #2. (C, D) miRCURY analysis of synthetic samples #1 and #2. A Poisson randomness test was used to evaluate if the fraction of homology related false positives at a given Ct detection threshold was significantly higher than expected. The expected fraction was defined as the number of potential false positives (the number of assays on a given platform targeting miRNAs not present in the investigated sample) that have sequence homology to miRNAs in the sample. As the synthetic samples #1 and #2 contain the same miRNAs (but in different concentrations) the expected fraction is the same for the two samples. For the TaqMan platform the expected fraction is 93 out of 565 (16%) miRNAs and for the miRCURY platform it is 102 out of 552 (18%). **p < 0.0001, *p < 0.05.
Jensen et al. BMC Genomics 2011 12:435 doi:10.1186/1471-2164-12-435