Open AccessSimple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol
Author(s) -
Viiu Paalme,
Mart Speek
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0018
Subject(s) - plasmid , rnase p , alkaline lysis , chromatography , lysis , precipitation , plasmid preparation , dna , chemistry , rna , lysis buffer , microbiology and biotechnology , biology , dna vaccination , gene , biochemistry , physics , meteorology , pbr322
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.