McEntee C M, Hudson A P
Department of Microbiology and Immunology, Medical College of Pennsylvania, Philadelphia 19129.
Anal Biochem. 1989 Feb 1;176(2):303-6. doi: 10.1016/0003-2697(89)90313-8.
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.
氯仿混合物中匀浆，可从多达100毫升未经原生质球化处理的酵母细胞（酿酒酵母）培养物中制备高质量RNA。这种制备方法的RNA产量与其他需要对细胞进行初步原生质球化处理的方法相当。回收RNA的质量和数量与所用酵母菌株和细胞生长培养基无关，该方法对处于对数生长期或稳定期的细胞同样有效。作为全细胞RNA混合物一部分回收的线粒体RNA可能会有轻微降解。对回收的RNA混合物中单个转录本的分析表明，用这种方法制备RNA时，不会对任何特定的单个转录本或任何一组转录本产生选择偏向。
