This document discusses the important impact of RNA quality on miRNA expression analysis, as well as the application of the Agilent 2100 Bioanalyzer and Agilent Small RNA Assay in RNA integrity assessment and miRNA analysis.
The document explains that RNA degradation significantly affects gene expression profiling. It highlights the Agilent 2100 Bioanalyzer as a gold standard tool for assessing RNA integrity using automated capillary electrophoresis with specific assays like the Eukaryote Total RNA Nano Assay and Small RNA Assay.
Studies presented in the text demonstrate that ongoing RNA degradation leads to an increase in quantification cycle (Cq) values, resulting in inaccurate expression data. The note concludes that maintaining high-quality RNA with a RIN value higher than 5 is essential for reliable results.
Samples with a RIN higher than 5 are recommended to ensure reliable expression analysis.
Degradation causes an overestimation of absolute miRNA amounts because unspecific smaller fragments enter the analytical range.