Fast and Accurate Method for Quantitating E. Coli Host-Cell DNA Contamination in Plasmid DNA Preparations
نویسندگان
چکیده
منابع مشابه
Fast and accurate method for quantitating E. coli host-cell DNA contamination in plasmid DNA preparations.
Plasmid DNA is being used successfully as a gene delivery vector in a variety of clinical applications. Similar to other pharmaceutical products for clinical use, the plasmid vectors must meet rigorous purity standards. One important contaminant is the DNA of the host cell used to produce the plasmids. We have developed a new method to accurately quantitate E. coli host-cell DNA in plasmid prep...
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Several methods have been reported for the small scale preparation of plasmid DNA for sequencing (1-9), but little attention has been given to the effect of the E. coli host strain on the quality of the sequence obtained from double-stranded DNA. It has been observed that endonuclease A (endA) E.coli strains may produce nicked plasmid DNA resulting in 'shadow' bands on sequencing gels (1, 8), b...
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Background: Existence of bacterial host cell DNA contamination in biopharmaceutical products is a potential risk factor for patients receiving these drugs. Hence, the quantity of contamination must be controlled under the regulatory standards. Although different methods such as hybridization assays have been employed to determine DNA impurities, these methods are labor, intensive and rather exp...
متن کاملRelevance of Endotoxin Contamination in Plasmid DNA
Endotoxins are known to be frequent contaminants in plasmid DNA extracted from bacteria. While many typical downstream applications such as sequencing and cloning are not affected by the presence of these molecules, endotoxins are of great concern to researchers in the pharmaceutical and biotechnology industries, since they may serve as potent inhibitors to more sensitive downstream application...
متن کاملEffects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism
BACKGROUND Although understanding of physiological interactions between plasmid DNA and its host is important for vector design and host optimization in many biotechnological applications, to our knowledge, global studies on plasmid-host interactions have not been performed to date even for well-characterized plasmids. RESULTS Escherichia coli cells, either devoid of plasmid DNA or bearing pl...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1999
ISSN: 0736-6205,1940-9818
DOI: 10.2144/99263rr03