DNA extraction method for screening yeast clones by PCR
DNA extraction method for screening yeast clones by PCR
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DOI：
10.2144/00284st02
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发表时间：
2000-04-01
期刊：
影响因子：
2.7
通讯作者：
Nishizawa, Y
中科院分区：
文献类型：
--
作者：
Akada, R;Murakane, T;Nishizawa, Y
本文介绍了一种分离酵母DNA的简单方法，该方法适用于作为PCR扩增的模板。仅用十二烷基硫酸钠处理就足以从酵母细胞中提取染色体DNA。将酵母菌落的细胞悬浮在小体积(约20亩L)的0.25%十二烷基硫酸钠溶液中，大力混合，离心。稀释后的上清液可直接作为模板，得到最终的PCR混合液中nn-SDS浓度小于0.01%。
A simple procedure for isolating yeast DNA suitable for use ns a template for PCR amplification is described. SDS treatment alone is sufficient for extraction of chromosomal DNA from yeast cells. Cells of a yeast colony are suspended in a small volume (about 20 mu L) of a 0.25% SDS solution, mixed vigorously and centrifuged. The supernatant cart be directly used as a template after dilution to give nn SDS concentration of less than 0.01% in the final PCR mixture.