A rapid and versatile method for the isolation of total RNA from the filamentous fungus Trichoderma sp.

RNA isolation is essential to study gene expression at the molecular level. However, RNA isolation is especially difficult in organisms such as filamentous fungi which contain compounds that bind and/or co-precipitate with RNA. No commercial kit is yet available for the isolation of high-quality RNA from these organisms. Furthermore, because of the large amounts of polysaccharides, the common protocols for RNA isolation usually result in poor yields when applied to Trichoderma. A simple and fast protocol for the isolation of RNA with high yields that effectively remove contaminating substances is introduced here. The protocol is based on the SDS/phenol cellular sample treatment method, with the following modifications: a phenol/chloroform extraction step for removal of proteins, DNA, and polysaccharides by precipitation without β-mercaptoethanol and liquid nitrogen. This procedure may be useful for the isolation of RNA from other filamentous fungi and may serve as an important tool for the molecular analysis of these organisms. [ABSTRACT FROM AUTHOR]