RNAintegriteit

RNA integrity refers to the state of intactness of RNA molecules within a sample. It is a crucial factor in the success of many molecular biology applications, particularly those involving RNA analysis, such as gene expression profiling, RNA sequencing, and quantitative reverse transcription polymerase chain reaction (RT-qPCR). Degraded RNA, where the phosphodiester bonds have been broken, can lead to inaccurate and unreliable experimental results.

The integrity of RNA is commonly assessed using techniques like gel electrophoresis or automated capillary electrophoresis. These methods separate RNA molecules by size and allow for the visualization of ribosomal RNA (rRNA) bands, which are typically abundant and relatively stable. A key metric used to quantify RNA integrity is the RNA Integrity Number (RIN). The RIN is a standardized scale from 1 to 10, where 1 represents completely degraded RNA and 10 represents intact, high-quality RNA. A higher RIN value indicates better RNA integrity.

Factors that can compromise RNA integrity include the presence of RNases (ribonucleases), which are enzymes that degrade RNA, as well as improper sample collection, storage, and handling procedures. Researchers often employ specific protocols and reagents to inhibit RNase activity and preserve RNA integrity throughout the experimental workflow. Maintaining high RNA integrity is essential for obtaining meaningful and reproducible biological insights from RNA-based studies.