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Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol.
Biotechniques ( IF 2.2 ) Pub Date : 2021-08-16 , DOI: 10.2144/btn-2021-0018
Viiu Paalme 1 , Mart Speek 1
Affiliation  

We present a modified alkaline lysis method for purification of plasmid DNA (pDNA) from bacterial extract using fractional precipitation with isopropanol (FPI). This method includes two successive precipitations with 0.33 and 0.36 volumes of isopropanol and separates pDNA from total RNA and most of the lipopolysaccharides. Using different quality control tests, we demonstrate that plasmids purified with FPI show superior quality compared to plasmids prepared with commercial kits based on spin-column chromatography.

中文翻译:

通过异丙醇分级沉淀,简单、廉价且无 RNase 纯化质粒 DNA。

我们提出了一种改进的碱裂解方法,用于使用异丙醇 (FPI) 分级沉淀从细菌提取物中纯化质粒 DNA (pDNA)。该方法包括使用 0.33 和 0.36 体积的异丙醇进行两次连续沉淀,并将 pDNA 从总 RNA 和大部分脂多糖中分离出来。我们使用不同的质量控制测试证明,与使用基于旋转柱色谱法的商业试剂盒制备的质粒相比,使用 FPI 纯化的质粒显示出更高的质量。
更新日期:2021-08-16
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