Preparation of Total RNA

RNA analysis is central to a detailed understanding of gene expression. The expression of many nuclear genes from a wide range of organisms has been studied at the protein level using reporter genes (lacZ , for example), and changes in expression levels in response to specific stimuli are frequently presumed to be mediated at the transcriptional level. However, there are an increasing number of examples of fungal and vertebrate genes that are post-transcriptionally regulated. It is therefore prudent to investigate expression at the RNA and protein levels. This chapter will discuss the isolation of total RNA from yeast. The reader should refer to ref. (1 ) for a detailed description of the methods for preparation of ribosomal and small RNAs from yeast.