Transformation of Schizosaccharomyces pombe: Electroporation Procedure.
Transformation of Schizosaccharomyces pombe: Electroporation Procedure.
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DOI：
10.1101/pdb.prot090951
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发表时间：
2016-04-01
影响因子：
--
通讯作者：
Carr, Antony M
中科院分区：
文献类型：
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作者：
Murray, Johanne M;Watson, Adam T;Carr, Antony M
用DNA转化粟酒裂殖酵母需要调节细胞以促进DNA摄取，然后在选择和维持质粒或整合事件的条件下进行细胞生长。三种主要的方法是电穿孔、用锂阳离子处理和原生质体转化。该方案描述了通过电穿孔的转化。它涉及预处理S。用二硫苏糖醇（DTT）处理粟酒酵母细胞，一旦施加电脉冲，其增加转化效率。
Transformation ofSchizosaccharomyces pombewith DNA requires the conditioning of cells to promote DNA uptake followed by cell growth under conditions that select and maintain the plasmid or integration event. The three main methodologies are electroporation, treatment with lithium cations, and transformation of protoplasts. This protocol describes transformation by electroporation. It involves pretreatingS. pombecells with dithiothreitol (DTT), which increases the transformation efficiency once the electric pulse is applied.