| Autor: |
A. Rita Silva-Santos, Alexandra Wagner, Duarte M. F. Prazeres, Ana Azevedo, Sophie Gierak, Sara A.S.L. Rosa |
| Rok vydání: |
2020 |
| Předmět: |
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| Zdroj: |
Methods in Molecular Biology ISBN: 9781071608715 |
| DOI: |
10.1007/978-1-0716-0872-2_8 |
| Popis: |
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%). |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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