This application note describes the use of the Agilent 2100 bioanalyzer for stringent RNA quality control and the advantages of this method for high-resolution analysis of small and very small RNA samples.
The document explains that successful RNA technology relies on high-quality RNA samples. It introduces the Agilent 2100 bioanalyzer as a chip-based system using pre-packed kits to reduce manual intervention and improve data accuracy compared to traditional gel electrophoresis.
A key focus of this note is the importance of heating RNA samples prior to electrophoresis. The text details how heating destabilizes base-pairing in fragmented rRNAs and aggregates, preventing misleading results where broken fragments mimic intact molecules. It provides examples from human, fish, and plant tissues.
Only about 25 ng of total RNA are required for analysis.
RNA samples should be incubated for 2 minutes at 70 °C in thin-wall 0.5 mL PCR tubes.
The migration difference is about 5.6 seconds for heated human RNA samples.