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Efficient Recovery of Ultrapure Plasmid DNA

Nicole Passaretti, Vincent Prezioso, PhD, and Susanne Wagner
Nicole Passaretti, Vincent Prezioso, PhDBrinkmann Instruments, Inc., Westbury, NY
Susanne WagnerEppendorfNethelerHinz GmbH, Hamburg, Germany Plasmids are naturally occurring, extrachromosomal DNA in a bacterial cell, which can replicate independently but (usually) do not integrate into the host chromosome. Those used for molecular cloning have been artificially created from various natural plasmids. Plasmids have become the cornerstone of modern molecular biology research, making nucleic acid purification a standard application in most laboratories. It is with these standard methods that users are looking for high-quality and high quantity results obtainable with an easy-to-handle, reproducible method. Perfectprep Plasmid Kits easily meet these essential demands. These ready-to-use kits were designed to suit the throughput and DNA quantities required in the lab: from single purification in a spin column (ranging in size from Mini to XL) to the simultaneous isolation of 96 plasmid samples.

Handling

The general process of plasmid recovery and purification is optimized by the new purification kits. An improved alkaline lysis step is used for plasmid isolation, which enables efficient separation of the plasmid DNA from chromosomal DNA.

Fig. 1: Restriction digestion of 40 individual plasmids with EcoRI (Roche Molecular Biochemicals). Vector pVL1392; length: 9.6 kb; bacteria strain: XL-1 Blue; medium 2x YT. The plasmid DNA was separated using the Perfectprep Plasmid
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