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用于通过聚合酶链反应筛选酵母克隆的DNA提取方法。

DNA extraction method for screening yeast clones by PCR.

作者信息

Akada R, Murakane T, Nishizawa Y

机构信息

Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Ube, Japan.

出版信息

Biotechniques. 2000 Apr;28(4):668-70, 672, 674. doi: 10.2144/00284st02.

DOI:10.2144/00284st02
PMID:10769744
Abstract

A simple procedure for isolating yeast DNA suitable for use as 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 microL) of a 0.25% SDS solution, mixed vigorously and centrifuged. The supernatant can be directly used as a template after dilution to give an SDS concentration of less than 0.01% in the final PCR mixture.

摘要

本文描述了一种简单的用于分离适合作为PCR扩增模板的酵母DNA的方法。单独使用SDS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于少量(约20微升)0.25%的SDS溶液中,剧烈混合并离心。上清液在稀释后可直接用作模板,以使最终PCR混合物中的SDS浓度低于0.01%。

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