As for DNA, quantification and normalization of RNA is traditionally done by measuring the absorbance at 260nm (A260). However, for impure RNA samples, the A260 read-out can lead to an overestimation of the RNA concentration due to UV-absorbing contaminants.
Alternative methods, such as the RiboGreen® assay, based on fluorescence enhancement upon binding with RNA, have been developed to address this issue. While offering improved specificity and sensitivity, such assays are less convenient, requiring multiple pipetting steps and running additional standards. Moreover, RiboGreen® assay is much less accurate than PicoGreen® assay for DNA samples.
Secondly A260/A280 and A260/A230 ratios are commonly used to quickly assess the purity of samples. Unfortunately those ratios are insensitive purity indicators and not reliable enough. Components as phenol or guanidinium salts used in extraction buffers can be copurified with the RNA. Such contaminants can drastically affect the success and accuracy of downstream applications.
The DropSense96 platform combined with cDrop software provides a tool for both specific RNA quantification, purity as well as degradation information.
For more details on how the DropSense96 and the cDrop software can help you regarding your main application, please see other sections in the left menu.
This technology is ideal to help with:
- Specific quantification of RNA
- QC of RNA samples prior storage, NGS, RT-PCR…
- Assessment of RNA degradation