Automated Purification of Transfection-Grade Plasmid DNA Using Wizard MagneSil Tfx System |
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Authors: | Douglas White Donald Creswell Elaine Schenborn Steve Krueger |
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Affiliation: | Available online 19 February 2004. |
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Abstract: | We describe a reagent system and robotic protocol for the isolation of highly purified plasmid DNA from cultured cells. The method is based on the Wizard® MagneSil™ Plasmid Purification System, which purifies sequencing-grade plasmid DNA. Two modifications to the sequencing-grade system were made to create the Wizard MagneSil Tfx™ System. This system provides lower endotoxin and other contaminant levels, giving higher quality plasmid for transfection applications. The Wizard MagneSil Tfx™ System uses MagneSil™ Paramagnetic Particles (PMPs) to clear lysate and bind plasmid, eliminating the need for filtration devices. The endotoxin removal step uses MagneSil™ (PMPs) and a guanidine/isopropanol wash to remove RNA and protein. One 96-well plate may be processed in 45 minutes on the Beckman Biomek® FX robotic workstation. We provide data showing DNA yield, contaminant levels, and transfection efficiency for 5 commonly used cell lines. Comparisons with other systems are also shown. |
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Keywords: | Author Keywords: AUTOMATED TRANSFECTION PLASMID MAGNESIL PARAMAGNETIC ENDOTOXIN |
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