Next generations sequencing (NGS) has become an important tool in biomedical research. The Primer ID approach combined with the MiSeq platform overcomes the limitation of PCR errors and reveals the true sampling depth of population sequencing, making it an ideal tool to study mutagenic effects of potential broad-spectrum antivirals on RNA viruses. In this report we describe a protocol using Primer ID sequencing to study the mutations induced by antivirals in a coronavirus genome from an in vitro cell culture model and an in vivo mouse model. Viral RNA or total lung tissue RNA is tagged with Primer ID-containing cDNA primers during the initial reverse transcription step, followed by two rounds of PCR to amplify viral sequences and incorporate sequencing adaptors. Purified and pooled

Most bacteria in natural ecosystems form biofilms-a bacterial community, surrounded by a polymer matrix that consists mostly of exopolysaccharides, proteins, and nucleic acids. Extracellular RNA as a matrix component is involved in biofilm formation-the fact that was confirmed by direct detection of extracellular RNA in the biofilm matrix, and by an interruption of the biofilm's structure with RNases. Number of protocols describing isolation of RNA from biofilm matrix are limited and usually involve uncommon equipment and reagents. Here we describe simple method for extracellular RNA isolation from biofilm matrix using basic laboratory reagent and equipment. Key steps of the protocol include separation of matrix and bacterial cells with high ionic solution of NaCl, RNA precipitation with