RAKE microRNA

MicroRNAs (miRNAs) are small regulatory RNAs, which serve fundamental biological roles across eukaryotic species. We describe a novel method for high throughput miRNA detection. The technique is termed RNA-primed, Array-based, Klenow Enzyme (RAKE) assay, because it involves on-slide application of the Klenow fragment of DNA polymerase to extend unmodified miRNAs hybridized to immobilized DNA probes. We used RAKE to study human cell lines and brain tumors. We show that the RAKE assay is sensitive and specific for miRNAs and is ideally suited for rapid expression profiling of all known miRNAs. RAKE offers unique advantages for specificity over Northern blots or other microarray-based expression profiling platforms. Furthermore, we demonstrate that miRNAs can be isolated and profiled from formalinfixed paraffin-embedded tissue, which opens up new opportunities for analyses of small RNAs from archival human tissue. The RAKE assay is theoretically versatile and may be used for other applications, such as viral gene profiling. Keywords = microRNA Keywords: ordered