Quantitation of plasmid DNA in aqueous two-phase systems by fluorescence analysis
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^sup -1^. Two polyethylene...
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Veröffentlicht in: | Biotechnology letters 2000-07, Vol.22 (13), p.1101-1104 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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^sup -1^. Two polyethyleneglycol (PEG)/salt systems were studied, PEG 600/K^sub 2^HPO^sub 4^ and PEG 300/K^sub 2^HPO^sub 4^. 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.[PUBLICATION ABSTRACT] |
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ISSN: | 0141-5492 1573-6776 |
DOI: | 10.1023/A:1005610113677 |