Hofmeister series salts enhance purification of plasmid DNA by non-ionic detergents.
Hofmeister series salts enhance purification of plasmid DNA by non-ionic detergents.
复制标题
Hofmeister 系列盐可增强非离子去污剂对质粒 DNA 的纯化。
DOI：
10.1002/bit.23116
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发表时间：
2011
影响因子：
3.8
通讯作者：
Brunelli,Luca
中科院分区：
文献类型：
--
作者：
Lezin,George;Kuehn,MichaelR;Brunelli,Luca
离子交换色谱是用于质粒DNA纯化的标准技术，这是一种基本的分子生物学程序。非离子去污剂（NID）已用于质粒DNA纯化，但尚不清楚霍夫迈斯特系列盐（HSS）是否会改变特定NID的溶解度和相分离特性，从而增强质粒DNA纯化。在放大NID介导的质粒DNA分离后，我们确定HSS溶液中的NID最大限度地减少了蛋白质对质粒DNA的污染。此外，大规模NID/HSS溶液比Qiagen离子交换柱更有效地消除了质粒DNA的脂多糖（LPS）污染。与碱分离/柱纯化相比，大规模NID分离/NID纯化产生了更高产量的高质量DNA。这项工作表征了HSS如何增强NID介导的质粒DNA纯化，并证明NID相变对于从质粒DNA中去除LPS不是必需的。特定的NID（如IGEPAL CA-520）可用于快速、廉价和有效的基于实验室的大规模质粒DNA纯化，优于基于Qiagen的柱程序。Biotechnol. Bioeng. 2011; 108：1872-1882.© 2011 Wiley Periodicals，Inc.
Ion‐exchange chromatography is the standard technique used for plasmid DNA purification, an essential molecular biology procedure. Non‐ionic detergents (NIDs) have been used for plasmid DNA purification, but it is unclear whether Hofmeister series salts (HSS) change the solubility and phase separation properties of specific NIDs, enhancing plasmid DNA purification. After scaling‐up NID‐mediated plasmid DNA isolation, we established that NIDs in HSS solutions minimize plasmid DNA contamination with protein. In addition, large‐scale NID/HSS solutions eliminated lipopolysaccharides (LPS) contamination of plasmid DNA more effectively than Qiagen ion‐exchange columns. Large‐scale NID isolation/NID purification generated increased yields of high‐quality DNA compared to alkali isolation/column purification. This work characterizes how HSS enhance NID‐mediated plasmid DNA purification, and demonstrates that NID phase transition is not necessary for LPS removal from plasmid DNA. Specific NIDs such as IGEPAL CA‐520 can be utilized for rapid, inexpensive, and efficient laboratory‐based large‐scale plasmid DNA purification, outperforming Qiagen‐based column procedures. Biotechnol. Bioeng. 2011; 108:1872–1882. © 2011 Wiley Periodicals, Inc.