RNA is the key component for all kinds of therapeutic and analytical applications. However, contaminants show up in RNA preps no matter the RNA source and these contaminants can interfere with anything from gene expression assays to creating RNA-LNPs.
Proteins and genomic DNA often show up in RNA extracted from tissue, cells, bacteria or plasma, because separating the phases during extraction is a tricky job. Improper washing of the RNA can also leave behind reagent residues. Protein and solvent contaminants can inhibit DNases and lower the amount of recovered RNA, while gDNA can interfere with RT-PCR – as a result, there’ll be increased noise in downstream applications like RNA-seq.
Residual DNA templates are a critical process-related impurity of in vitro RNA transcription. When making RNA-LNPs, this DNA can be incorporated into the nanoparticles alongside the RNA and increase oncogenic, infective, or immunomodulatory risks.
Traditional RNA quality control by UV/Vis spectroscopy involves checking if the A260/A280 ratio is ~2.0, indicating pure RNA. However, a ratio below 2.0 says nothing about the identity of the contaminants and it can leave you in the dark about the actual RNA quantity. This is where Lunatic and its Unmix applications flex their muscles.