Preparation of RNA from unspheroplasted yeast cells (Saccharomyces cerevisiae).
High-quality RNA can be prepared from up to 100-ml culture volumes of unspheroplasted yeast cells (Saccharomyces cerevisiae) via homogenization in high-temperature phenol:chloroform mixtures. The yield of RNA from this preparative method is equivalent to those of other methods requiring preliminary spheroplasting of cells. Quality and quantity of recovered RNA are independent of yeast strain and cell growth medium used, and the method works equally well on cells in either log phase growth or in stationary phase. Mitochondrial RNAs recovered as part of whole cell RNA mixtures may be slightly degraded. Analyses of individual transcripts in the recovered RNA mixtures suggest that there is no selection for or against any specific single transcript or any group of transcripts when RNA is prepared by this method.
McEntee
C M
CM
Department of Microbiology and Immunology, Medical College of Pennsylvania, Philadelphia 19129.
Hudson
A P
AP
D015152:Blotting, Northern; D004587:Electrophoresis, Agar Gel; D005800:Genes, Fungal; D006358:Hot Temperature; D019800:Phenol; D010636:Phenols / Q000494:pharmacology; D012331:RNA, Fungal / Q000032:analysis / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013104:Spheroplasts / Q000032:analysis*
D010636:Phenols; D012331:RNA, Fungal; D019800:Phenol