In the automated workflow, viral RNA binds specifically to the magnetic beads while contaminants such as divalent cations and proteins, are completely removed in three efficient wash steps. High-quality RNA is eluted in a special RNase-free buffer, ready for direct use or safe storage. The purified RNA is free of proteins, nucleases, and other contaminants or inhibitors and can be used immediately in demanding downstream applications, including PCR, qPCR, dPCR and next-generation sequencing. Protocols are available for use on the Thermo Scientific KingFisher Flex. This kit can also be used with many other automated handling systems.
Carrier RNA serves two purposes: First, it enhances binding of viral nucleic acids to the magnetic beads, especially if there are very few target molecules in the sample. Secondly, the addition of large amounts of carrier RNA reduces the chance of viral RNA degradation in the rare event that RNase molecules escape denaturation. Not adding carrier RNA to Buffer ACL may lead to reduced viral RNA recovery.