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Primary Purification of Plasmid DNA Using Differential Isopropanol Precipitation
- Source :
- Methods in molecular biology (Clifton, N.J.). 2197
- Publication Year :
- 2020
-
Abstract
- A method for the intermediate recovery of plasmid DNA (pDNA) from alkaline lysates is described that comprises differential isopropanol precipitation steps. In a first low-cut precipitation, a smaller amount of isopropanol (20% v/v) is used so that only high molecular weight RNA precipitates. After solid liquid separation, a high-cut precipitation is performed by bringing isopropanol concentration to 70% v/v to precipitate pDNA. Tests made with lysates show that the differential precipitation increases purity threefold compared to the conventional one-step precipitation at 70% v/v without affecting pDNA recovery (80%).
Details
- ISSN :
- 19406029
- Volume :
- 2197
- Database :
- OpenAIRE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Accession number :
- edsair.pmid..........e852c80062dc7849c9ca8f0ea4e4b348