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Paalme V, Speek M. Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol. Biotechniques. 2021;71(3):490-494doi: 10.2144/btn-2021-0018.
Paalme, V., & Speek, M. (2021). Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol. BioTechniques, 71(3), 490-494. https://doi.org/10.2144/btn-2021-0018
Paalme, Viiu, and Speek, Mart. "Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol." BioTechniques vol. 71,3 (2021): 490-494. doi: https://doi.org/10.2144/btn-2021-0018
Paalme V, Speek M. Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol. Biotechniques. 2021 Sep;71(3):490-494. doi: 10.2144/btn-2021-0018. Epub 2021 Aug 16. PMID: 34392702.
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Biotechniques. 2021 Sep;71(3):490-494. doi: 10.2144/btn-2021-0018. Epub 2021 Aug 16.

Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol.

BioTechniques

Viiu Paalme, Mart Speek

Affiliations

  1. Department of Chemistry & Biotechnology, Tallinn University of Technology, Akadeemia Tee 15, Tallinn, 12618, Estonia.

PMID: 34392702 DOI: 10.2144/btn-2021-0018

Abstract

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.

Keywords: RNase; alkaline lysis; fractional precipitation; isopropanol; lipopolysaccharides; plasmid DNA

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