Hoffman C S
Boston College, Chestnut Hill, Massachusetts, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 13:Unit13.11. doi: 10.1002/0471142727.mb1311s39.
Molecular studies in yeast often require the isolation of both plasmid and chromosomal yeast DNA. Plasmid DNA is used in the transformation of E. coli, whereas chromosomal DNA is used for Southern hybridization analysis, in vitro amplification by the polymerase chain reaction (PCR), or cloning of integrated plasmids. This unit presents two variations of the "smash and grab" protocol that produce suitable DNA for all these applications. These protocols work for both Saccharomyces cerevisiae and Schizosaccharomyces pombe.
酵母中的分子研究通常需要分离质粒和染色体酵母DNA。质粒DNA用于大肠杆菌的转化,而染色体DNA用于Southern杂交分析、通过聚合酶链反应(PCR)进行体外扩增或整合质粒的克隆。本单元介绍了“粉碎与抓取”方案的两种变体,它们能产生适用于所有这些应用的DNA。这些方案适用于酿酒酵母和粟酒裂殖酵母。