This Application Note investigates the various techniques for RNA quantitation, including a comparison of the Agilent 2100 bioanalyzer with UV spectrometry and the Ribogreen® quantitation system to access the linear dynamic range, reproducibility, and the effects of contaminants for each quantitation method.
This document investigates techniques for RNA quantitation, specifically comparing the Agilent 2100 bioanalyzer with the RNA 6000 Nano assay against UV spectrometry and Ribogreen analysis. The study evaluates these methods based on their linear dynamic range, reproducibility, and accuracy in the presence of common contaminants such as genomic DNA and phenol.
The analysis covers the performance of the Agilent 2100 bioanalyzer using the RNA 6000 Nano LabChip kit, which utilizes microfluidic technology and laser excitation. Results indicate that while UV analysis is stable, it is susceptible to contaminants. Ribogreen offers high sensitivity but can be affected by contaminants and phenol. The Agilent 2100 bioanalyzer is highlighted for its ability to accurately quantify RNA in the presence of contaminants while consuming only 1 µL of sample.
| Linear dynamic range | 25 ng/ul to 500 ng/ul |
|---|---|
| Linearity R2 (RNA 6000 Nano) | 0.9976 |
| Linearity R2 (UV analysis) | 0.9984 |
| Linearity R2 (Ribogreen) | 0.9977 |
The assay has a specified linear dynamic range of quantitation from 25 ng/ul to 500 ng/ul.
The 2100 bioanalyzer can accurately quantitate in the presence of contaminants, whereas UV analysis is impacted by contaminants like genomic DNA or phenol.