Total RNA isolation from recalcitrant yeast cells.

Conventional methods of RNA isolation are not suitable for yeast cells from stationary phase and fermentation broth. Methods specially reported for such cells are cumbersome and do not lend themselves for use with large number of samples. Here we report a facile method of RNA isolation from such recalcitrant yeast cells. The entire procedure is performed in microcentrifuge tubes and, thus, is ideal for faster processing of multiple samples. The method consistently gives high quality and quantity of RNA, which was found to be suitable for downstream applications such as quantitative real-time polymerase chain reaction. Besides Saccharomyces cerevisiae, the method was found to work equally well with other yeast species; thus, it is likely to have wider applicability.