A PCR-based platform for microRNA expression profiling studies

MicroRNAs (miRNAs) are newly identified small noncoding RNA molecules that play important regulatory roles in many biological processes. Changes in miRNA expression are associated with a variety of normal physiological processes as well as with human diseases including cancer. Thus, one important way to characterize miRNA function is to identify changes in miRNA expression profiles. To date, microarrays are the most widely used technique for miRNA expression profiling analysis. However, there are major limitations when microarrays are applied to miRNA profiling studies. Here we report a new PCR-based assay platform for miRNA expression profiling analysis. The primers associated with these miRNA assays were designed using a novel bioinformatics algorithm that has incorporated many primer selection features for improved assay specificity, sensitivity, and homogeneity. The primer design algorithm has been extensively validated experimentally and has been shown to have robust performance for the measurement of miRNA expression in a variety of biological samples at a low cost.