RNAprotect Bacteria Reagent (2x100ml)

From QIAGEN, this reagent is intended for research use only and is designed for in vivo stabilization of total RNA in bacterial cells, preventing both enzymatic RNA degradation and changes in gene expression that normally occur during harvesting and handling. The formulation provides immediate stabilization of the gene expression profile, so RNA isolated afterward reflects the true in vivo state. It is suitable for both Gram-positive bacteria (such as Staphylococcus aureus and Mycobacterium avium) and Gram-negative bacteria (such as Escherichia coli and Salmonella typhimurium), and is compatible with both minimal and complex culture media.

Two volumes of the reagent are added directly to one volume of bacterial culture, providing immediate RNA stabilization. Alternatively, two volumes of reagent mixed with one volume of media or PBS can be applied to bacteria grown on solid media. After stabilization, bacterial cells are disrupted to release RNA, which is then purified using the RNeasy Mini Kit or RNeasy Midi Kit. The protocol options include enzymatic lysis with lysozyme or lysostaphin, proteinase K digestion, and mechanical disruption using the TissueLyser II with glass beads, allowing users to select an approach suited to their organism and starting material. Purified RNA is suitable for northern, dot and slot blotting; array analyses; quantitative real-time RT-PCR (such as QuantiTect technology); and other nucleic acid-based methods including NASBA and bDNA analyses.

The reagent should be stored at room temperature (15–25°C), under which conditions it is stable for at least 12 months.