Quantitation of plasmid DNA in aqueous two-phase systems by fluorescence analysis |
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| Authors: | Sofia C Ribeiro Gabriel A Monteiro Gaspar Martinho Joaquim MS Cabral Duarte MF Prazeres |
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| Institution: | (1) Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049-001 Lisboa, Portugal;(2) Centro de Química-Física Molecular, Instituto Superior Técnico, 1049-001 Lisboa, Portugal |
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| Abstract: | The development of aqueous two-phase systems for plasmid purification from Escherichia coli cell lysates requires a reliable DNA quantitation method. Plasmid DNA was quantified by fluorescence using PicoGreen nucleic acid stain. Linearity was obtained up to 40 ng plasmid ml–1. Two polyethyleneglycol (PEG)/salt systems were studied, PEG 600/K2HPO4 and PEG 300/K2HPO4. The average plasmid recovery was 41% in the bottom phase of the first system and 35% in the top phase of the second system. This method has proved to be simple and reproducible. |
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| Keywords: | aqueous two-phase systems fluorescence PicoGreen plasmid DNA |
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