This novel method for plasmid preparation in 96-well format provides highly pure, transfection-grade plasmid DNA using a simple bind-wash-elute procedure (see flowchart " QIAGEN PlasmidPlus 96 procedure").
The QIAGEN Plasmid Plus 96 Miniprep protocol starts with a modified alkaline lysis procedure. After neutralization, lysates are cleared using a TurboFilter 96 plate. A nonchaotropic binding buffer is added to the cleared lysate to optimize plasmid DNA binding to the membrane of the Plasmid Plus 96 plate. The unique binding buffer provides very specific binding conditions leading to DNA quality that is comparable to anion-exchange preps. A wash step with an endotoxin reduction buffer results in transfection-grade plasmid DNA suited for transfection into most cell lines (see figure " Highly efficient transfection into a sensitive cell lines"). The proprietary binding chemistry allows purification of up to 50 μg plasmid DNA per well from a 5 ml E. coli culture (see figure " High yields of plasmid DNA").
Convenient and flexible processing options
The procedure may be fully automated using the QIAGEN Plasmid Plus 96 BioRobot Kit on the BioRobot Universal System. Alternatively, the QIAvac 96 is used for lysate clearing using the TurboFilter 96 plate, and DNA binding and wash steps are performed using the Plasmid Plus 96 plate. Drying and elution steps may either be performed on the QIAvac 96 or using a compatible centrifuge. In addition, the complete procedure can be performed using a compatible centrifuge.