Rapid quantitation and monitoring of plasmid DNA using an ultrasensitive DNA-binding dye
| Autor: | Ronan D. O'Kennedy, Eli Keshavarz-Moore, N. Abidi, M. S. Levy, I. S. Noites |
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| Rok vydání: | 1999 |
| Předmět: |
Plasmid preparation
Lysis Chromatography food and beverages Bioengineering Biology medicine.disease_cause biology.organism_classification Applied Microbiology and Biotechnology Fluorescence Molecular biology chemistry.chemical_compound Plasmid chemistry medicine Fermentation Escherichia coli Bacteria DNA Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 66:195-201 |
| ISSN: | 1097-0290 0006-3592 |
| DOI: | 10.1002/(sici)1097-0290(1999)66:3<195::aid-bit8>3.0.co;2-6 |
| Popis: | A sensitive fluorescence-based method for monitoring plasmid DNA during production was investigated. This simple method of assaying for plasmid DNA allows rapid monitoring of plasmid yields from a recombinant Escherichia coli fed-batch fermentation. The assay has several advantages over traditional methods of plasmid DNA measurement. The fluorescent dye is highly specific and can measure total plasmid DNA concentration in about 5 min. The assay is sensitive over a wide range of plasmid concentrations of between 15 and 280 ng/mL, even in the presence of impurities that occur within alkaline lysate preparations. The technique can also be applied to monitoring fermentation and downstream purification steps. |
| Databáze: | OpenAIRE |
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